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Monsigny Michel


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tél : 02.38.25.55.57 - fax : 02.38.25.78.07

Publications

2010   Références trouvées : 1

Duverger E., Lamerant-Fayel N., Frison N. & Monsigny M.  (2010)

Carbohydrate-lectin interactions assayed by SPR

In "Surface Plasmon Resonance : Methods and Protocols" - De Mol N.J., Fisher M.J.E (Eds.) 627 : 157-178


2008   Références trouvées : 1

Grosse, S., Thevenot, G., Aron, Y., Duverger, E., Abdelkarim, M., Roche, A.C., Monsigny, M. & Fajac, I.  (2008)

In vivo gene delivery in the mouse lung with lactosylated polyethylenimine, questioning the relevance of in vitro experiments.

J. Control. Release 132, 105-112.


2007   Références trouvées : 2

Srinivas, O ; Larrieu, P ; Duverger, E ; Boccaccio, C ; Bousser, MT ; Monsigny, M ; Fonteneau, JF ; Jotereau, F ; Roche, AC  (2007)

Synthesis of glycocluster-tumor antigenic peptide conjugates for dendritic cell targeting

Bioconjugate Chemistry 18 1547-1554
The use of dendritic cells (DC) for the development of therapeutic cancer vaccines is attractive because of their unique ability to present tumor epitopes via the MHC class I pathway to induce cytotoxic CD8(+) T lymphocyte responses. C-Type membrane lectins, DC-SIGN and the mannose receptor (MR), present on the DC surface, recognize oligosaccharides containing mannose and/or fucose and mediate sugar-specific endocytosis of synthetic oligolysine-based glycoclusters. We therefore asked whether a glycotargeting approach could be used to induce uptake and presentation of tumor antigens by DC. To this end, we designed and synthesized glycocluster conjugates containing a CD8(+) epitope of the Melan-A/Mart-l melanoma antigen.

The use of dendritic cells (DC) for the development of therapeutic cancer vaccines is attractive because of their unique ability to present tumor epitopes via the MHC class I pathway to induce cytotoxic CD8(+) T lymphocyte responses. C-Type membrane lectins, DC-SIGN and the mannose receptor (MR), present on the DC surface, recognize oligosaccharides containing mannose and/or fucose and mediate sugar-specific endocytosis of synthetic oligolysine-based glycoclusters. We therefore asked whether a glycotargeting approach could be used to induce uptake and presentation of tumor antigens by DC. To this end, we designed and synthesized glycocluster conjugates containing a CD8(+) epitope of the Melan-A/Mart-l melanoma antigen.

Grosse, S ; Aron, Y ; Thevenot, G ; Monsigny, M ; Fajac, I  (2007)

Cytoskeletal involvement in the cellular trafficking of plasmid/PEI derivative complexes

Journal of Controlled Release 122 111-117
We have studied the cytoskeletal involvement in the cellular trafficking of complexes made with plasmid/PEI or plasmid/lactosylated PEI in cystic fibrosis airway epithelial cells (Sigma CFTE29o-cells). Complexes were incubated in the presence of cytoskeletal inhibitors, and the number of transfected cells was determined by flow cytometry. Complexes were also generated with fluorescein-labeled PEI derivatives and the cell fluorescence intensity was determined by flow cytometry.

We have studied the cytoskeletal involvement in the cellular trafficking of complexes made with plasmid/PEI or plasmid/lactosylated PEI in cystic fibrosis airway epithelial cells (Sigma CFTE29o-cells). Complexes were incubated in the presence of cytoskeletal inhibitors, and the number of transfected cells was determined by flow cytometry. Complexes were also generated with fluorescein-labeled PEI derivatives and the cell fluorescence intensity was determined by flow cytometry.


2006   Références trouvées : 1

Grosse, S ; Thevenot, G ; Monsigny, M ; Fajac, I  (2006)

Which mechanism for nuclear import of plasmid DNA complexed with polyethylenimine derivatives ?

Journal of Gene Medicine 8 (7) 845-851
Background : To investigate the nuclear import mechanism of plasmid/polyethylenimine (PEI) derivative complexes and the putative nuclear targeting of therapeutic genes by the use of oligosaccharides, we have studied the nuclear import of plasmid DNA complexed either with PEI or with lactosylated PEI (Lac-PEI) in cystic fibrosis human airway epithelial cells (Sigma CFTE290-cells).

Background : To investigate the nuclear import mechanism of plasmid/polyethylenimine (PEI) derivative complexes and the putative nuclear targeting of therapeutic genes by the use of oligosaccharides, we have studied the nuclear import of plasmid DNA complexed either with PEI or with lactosylated PEI (Lac-PEI) in cystic fibrosis human airway epithelial cells (Sigma CFTE290-cells).


2005   Références trouvées : 1

Grosse, S ; Aron, Y ; Thevenot, G ; Francois, D ; Monsigny, M ; Fajac, I  (2005)

Potocytosis and cellular exit of complexes as cellular pathways for gene delivery by polycations

Journal of Gene Medicine 7 (10) 1275-1286
Background : Although polycations are among the most efficient nonviral vectors for gene transfer, the gene expression they allow is still too low for in vivo applications. To engineer more potent polycationic vectors, the factors governing the intracellular trafficking of a plasmid complexed with current polycations need to be identified.

Background : Although polycations are among the most efficient nonviral vectors for gene transfer, the gene expression they allow is still too low for in vivo applications. To engineer more potent polycationic vectors, the factors governing the intracellular trafficking of a plasmid complexed with current polycations need to be identified.


2004   Références trouvées : 2

Grosse, S ; Aron, Y ; Honore, I ; Thevenot, G ; Danel, C ; Roche, AC ; Monsigny, M ; Fajac, I  (2004)

Lactosylated polyethylenimine for gene transfer into airway epithelial cells : role of the sugar moiety in cell delivery and intracellular trafficking of the complexes

Journal of Gene Medicine 6 (3) 345-356
Background As we have previously shown that lactosylated polyethylenimine (PEI) is the most efficient glycosylated PEI for gene transfer into human airway epithelial cells in primary culture, we have studied here the role of the lactose residue in the enhancement of gene transfer efficiency observed with lactosylated PEI as compared with unsubstituted PEI in immortalized (SigmaCFTE29o- cells) and primary human airway epithelial cells. Methods and results After three transfections of 1 h performed daily, 60% of SigmaCFTE29o- cells were transfected with lactosylated PEI, whereas 25% of cells were transfected with unsubstituted PEI (p

Background As we have previously shown that lactosylated polyethylenimine (PEI) is the most efficient glycosylated PEI for gene transfer into human airway epithelial cells in primary culture, we have studied here the role of the lactose residue in the enhancement of gene transfer efficiency observed with lactosylated PEI as compared with unsubstituted PEI in immortalized (SigmaCFTE29o- cells) and primary human airway epithelial cells. Methods and results After three transfections of 1 h performed daily, 60% of SigmaCFTE29o- cells were transfected with lactosylated PEI, whereas 25% of cells were transfected with unsubstituted PEI (p

Monsigny, M ; Rondanino, C ; Duverger, E ; Fajac, I ; Roche, AC  (2004)

Glyco-dependent nuclear import of glycoproteins, glycoplexes and glycosylated plasmids

Biochimica et Biophysica Acta-General Subjects 1673 (1-2) 94-103
This short review deals with some properties of nuclear sugar-binding proteins also called nuclear lectins, the sugar-dependent nuclear import of neoglycoproteins and the attempts of using this pathway to enhance the nuclear import of plasmids in order to hopefully increase the expression of transferred genes. (C) 2004 Elsevier B.V. All rights reserved.

This short review deals with some properties of nuclear sugar-binding proteins also called nuclear lectins, the sugar-dependent nuclear import of neoglycoproteins and the attempts of using this pathway to enhance the nuclear import of plasmids in order to hopefully increase the expression of transferred genes. (C) 2004 Elsevier B.V. All rights reserved.


2003   Références trouvées : 9

Rondanino, C ; Bousser, MT ; Monsigny, M ; Roche, AC  (2003)

Sugar-dependent nuclear import of glycosylated proteins in living cells

Glycobiology 13 (7) 509-519
The nuclear import of proteins larger than M-r 40,000 depends on the presence of a nuclear localization signal (NLS) corresponding either to a short peptide sequence or to defined sugars. The sugar-dependent nuclear import was previously evidenced by using glycosylated proteins (neoglycoproteins) introduced into the cytosol of cells either by electroporation or on digitonin-permeabilization and was shown to be distinct from the peptide NLS-mediated pathway. In this work, we used a microinjection approach to compare the two nuclear import pathways in intact living cells. The intracellular localization of fluorescent NLS-BSA or Glc-BSA injected into the cytosol was analyzed by confocal microscopy. Novel differences between the two mechanisms were evidenced. First, Glc-BSA migrated less efficiently into the nucleus than NLS-BSA because of a cytosolic retention.

The nuclear import of proteins larger than M-r 40,000 depends on the presence of a nuclear localization signal (NLS) corresponding either to a short peptide sequence or to defined sugars. The sugar-dependent nuclear import was previously evidenced by using glycosylated proteins (neoglycoproteins) introduced into the cytosol of cells either by electroporation or on digitonin-permeabilization and was shown to be distinct from the peptide NLS-mediated pathway. In this work, we used a microinjection approach to compare the two nuclear import pathways in intact living cells. The intracellular localization of fluorescent NLS-BSA or Glc-BSA injected into the cytosol was analyzed by confocal microscopy. Novel differences between the two mechanisms were evidenced. First, Glc-BSA migrated less efficiently into the nucleus than NLS-BSA because of a cytosolic retention.

Frison, N ; Taylor, ME ; Soilleux, E ; Bousser, MT ; Mayer, R ; Monsigny, M ; Drickamer, K ; Roche, AC  (2003)

Oligolysine-based oligosaccharide clusters - Selective recognition and endocytosis by the mannose receptor and dendritic cell-specific intercellular adhesion molecule 3 (ICAM-3)-grabbing nonintegrin

Journal of Biological Chemistry 278 (26) 23922-23929
Dendritic cells are potent antigen-presenting cells that express several membrane lectins, including the mannose receptor and DC-SIGN ( dendritic cell-specific ICAM-3-grabbing nonintegrin). To identify highly specific ligands for these dendritic cell receptors, oligosaccharides were converted into glycosynthons (Os-1) and were used to prepare oligolysine-based glycoclusters, Os-[Lys(Os)](n)-Ala-Cys-NH2. Clusters containing two to six dimannosides as well as clusters containing four or five pentasaccharides (Lewis(a) or Lewis(x)) or hexasaccharides (Lewis(b)) were synthesized.

Dendritic cells are potent antigen-presenting cells that express several membrane lectins, including the mannose receptor and DC-SIGN ( dendritic cell-specific ICAM-3-grabbing nonintegrin). To identify highly specific ligands for these dendritic cell receptors, oligosaccharides were converted into glycosynthons (Os-1) and were used to prepare oligolysine-based glycoclusters, Os-[Lys(Os)](n)-Ala-Cys-NH2. Clusters containing two to six dimannosides as well as clusters containing four or five pentasaccharides (Lewis(a) or Lewis(x)) or hexasaccharides (Lewis(b)) were synthesized.

Roche, AC ; Fajac, I ; Grosse, S ; Frison, N ; Rondanino, C ; Mayer, R ; Monsigny, M  (2003)

Glycofection : facilitated gene transfer by cationic glycopolymers

Cellular and Molecular Life Sciences 60 (2) 288-297
Mammalian cells express several types of lectins involved in intracellular trafficking, including endocytosis, interorganelle routing and putatively nuclear import. In order to enhance the gene transfer efficiency, glycosylated cationic polymers have been used as nonviral vectors. We developed a simple method to convert reducing saccharides into glycosynthons. Glycosynthons are used to synthesize cationic glycopolymers, called Glycofectins. Glycofectins interact with a plasmid to give a glycoplex, a compacted form of a polymer/DNA complex. The high glycoplex efficiency depends on the sugar involved in the uptake and in the intracellular trafficking of glycoplexes. The present paper deals with glycoplexes, with gene transfer into cystic fibrosis airway epithelial and gland serous cells, and with some of the problems that have to be solved before clinical trials.

Mammalian cells express several types of lectins involved in intracellular trafficking, including endocytosis, interorganelle routing and putatively nuclear import. In order to enhance the gene transfer efficiency, glycosylated cationic polymers have been used as nonviral vectors. We developed a simple method to convert reducing saccharides into glycosynthons. Glycosynthons are used to synthesize cationic glycopolymers, called Glycofectins. Glycofectins interact with a plasmid to give a glycoplex, a compacted form of a polymer/DNA complex. The high glycoplex efficiency depends on the sugar involved in the uptake and in the intracellular trafficking of glycoplexes. The present paper deals with glycoplexes, with gene transfer into cystic fibrosis airway epithelial and gland serous cells, and with some of the problems that have to be solved before clinical trials.

Frison, N ; Meunier, L ; Marceau, P ; Quetard, C ; Strecker, G ; Roche, AC ; Mayer, R ; Monsigny, M  (2003)

Chromatographic separation of Lewis a and Lewis x oligosaccharides as glycosynthons

Biochimie 85 (1-2) 47-51
Lewis a and Lewis x oligosaccharides Galbeta3(Fucalpha4)GlcNAcbeta3Galbeta4Glc and Galbeta4(Fucalpha3)GlcNAcbeta3Galbeta4Glc are easily isolated as a mixture from biological fluids, including human milk. However, because they behave almost identically in most chromatographic systems, it is difficult to have each of them as a pure compound. Incidentally, we found that they were easily separated by HPLC as glycosynthons [Galbeta3(Fucalpha4)GlcNAcbeta3Galbeta4Glc-Glp-betaAla-OBzl and Galbeta4(Fucalpha3)GlcNAcbeta3Galbeta4Glc-Glp-betaAla-OBzl] after substitution of the terminal reducing sugar by a short peptide (pyroglutamyl-betaalanyl-O-benzyl ester) in a one-pot two-step reaction (Carbohydr. Lett. 1 (1995) 269 ; Bioconjug. Chem. 9 (1998) 268).

Lewis a and Lewis x oligosaccharides Galbeta3(Fucalpha4)GlcNAcbeta3Galbeta4Glc and Galbeta4(Fucalpha3)GlcNAcbeta3Galbeta4Glc are easily isolated as a mixture from biological fluids, including human milk. However, because they behave almost identically in most chromatographic systems, it is difficult to have each of them as a pure compound. Incidentally, we found that they were easily separated by HPLC as glycosynthons [Galbeta3(Fucalpha4)GlcNAcbeta3Galbeta4Glc-Glp-betaAla-OBzl and Galbeta4(Fucalpha3)GlcNAcbeta3Galbeta4Glc-Glp-betaAla-OBzl] after substitution of the terminal reducing sugar by a short peptide (pyroglutamyl-betaalanyl-O-benzyl ester) in a one-pot two-step reaction (Carbohydr. Lett. 1 (1995) 269 ; Bioconjug. Chem. 9 (1998) 268).

Duverger, E ; Frison, N ; Roche, AC ; Monsigny, M  (2003)

Carbohydrate-lectin interactions assessed by surface plasmon resonance

Biochimie 85 (1-2) 167-179
The specificity, the strength, the kinetics and some thermodynamic parameters of sugar-protein interactions are easily assessed by surface plasmon resonance (SPR). This paper intends to present both theoretical and practical considerations. This includes : the principle of SPR, the analysis according to Langmuir and Scatchard, the problems linked either to mass transport limitation, to the heterogeneity of the immobilized ligand density or to the non-linearity due to cluster effects. The non-linearity may be taken into account by either one of two ways : the fractal or the Sips approaches that have been developed with the aim of linearizing the data.

The specificity, the strength, the kinetics and some thermodynamic parameters of sugar-protein interactions are easily assessed by surface plasmon resonance (SPR). This paper intends to present both theoretical and practical considerations. This includes : the principle of SPR, the analysis according to Langmuir and Scatchard, the problems linked either to mass transport limitation, to the heterogeneity of the immobilized ligand density or to the non-linearity due to cluster effects. The non-linearity may be taken into account by either one of two ways : the fractal or the Sips approaches that have been developed with the aim of linearizing the data.

Fajac, I ; Thevenot, G ; Bedouet, L ; Danel, C ; Riquet, M ; Merten, M ; Figarella, C ; Ava-Santucci, JD ; Monsigny, M ; Briand, P  (2003)

Uptake of plasmid/glycosylated polymer complexes and gene transfer efficiency in differentiated airway epithelial cells

Journal of Gene Medicine 5 (1) 38-48
Background : We have studied gene transfer efficiency of glycosylated polylysines and glycosylated polyethylenimines as vectors in immortalized differentiated airway gland serous cells and primary cultures of human airway surface epithelial cells.

Background : We have studied gene transfer efficiency of glycosylated polylysines and glycosylated polyethylenimines as vectors in immortalized differentiated airway gland serous cells and primary cultures of human airway surface epithelial cells.

Grosse, S ; Honore, I ; Favatier, F ; Frison, N ; Monsigny, M ; Fajac, I  (2003)

Transcription and expression of plasmid DNA : Modulation by polyethylenimine/plasmid DNA complex formation

Molecular Therapy 7 (5) S374-S374 Part 2

Frison, N ; Taylor, ME ; Soilleux, E ; Bousser, MT ; Mayer, R ; Monsigny, M ; Drickamer, K ; Roche, AC  (2003)

Oligolysine-based oligosaccharide clusters. Selective recognition and endocytosis by the mannose receptor and dendritic cell-specific intercellular adhesion molecule 3 (ICAM-3)-grabbing nonintegrin

Journal of Biological Chemistry 278 (41) 40415-40415

Fajac, I ; Monsigny, M  (2003)

Glycotargeting of DNA

Journal of Gene Medicine 5 (3) S8-S8


2002   Références trouvées : 5

de Diesbach, P ; N’Kuli, F ; Berens, C ; Sonveaux, E ; Monsigny, M ; Roche, AC ; Courtoy, PJ  (2002)

Receptor-mediated endocytosis of phosphodiester oligonucleotides in the HepG2 cell line : evidence for non-conventional intracellular trafficking

Nucleic Acids Research 30 (7) 1512-1521
Having identified an oligonucleotide (ON) receptor in the HepG2 cell line, we have re-examined here the kinetics of ON uptake, subcellular distribution and intracellular localisation in these cells, at concentrations relevant for the study of a receptor-dependent process. Kinetic parameters of ON endocytosis were comparable with those of the receptor-mediated endocytosis tracer, transferrin (uptake equilibrium, saturation with concentration, specific competition and rapid efflux) and were clearly distinct from those of fluid-phase endocytosis.

Having identified an oligonucleotide (ON) receptor in the HepG2 cell line, we have re-examined here the kinetics of ON uptake, subcellular distribution and intracellular localisation in these cells, at concentrations relevant for the study of a receptor-dependent process. Kinetic parameters of ON endocytosis were comparable with those of the receptor-mediated endocytosis tracer, transferrin (uptake equilibrium, saturation with concentration, specific competition and rapid efflux) and were clearly distinct from those of fluid-phase endocytosis.

Frison, N ; Marceau, P ; Roche, AC ; Monsigny, M ; Mayer, R  (2002)

Oligolysine-based saccharide clusters : synthesis and specificity

Biochemical Journal 368 111-119 Part 1
In search of specific and highly selective sugar clusters for cell receptors, such as membrane lectins, various disaccharides were coupled to small peptide cores through an amide bond. In a first step, the reducing disaccharides, i.e. lactose and three different dimannoses, were converted into glycosyl-pyroglutamyl-ß-alanine derivatives. The free carboxylic group of these conjugates was then coupled to the a and epsilon amino groups of the core peptide (Lys(n)-Ala-Cys-NH2) with n = 1 to 5, with complete substitution leading to homogeneous glycoclusters. The thiol group of the cysteine residue was used to tag the glycosylated oligolysines upon reaction with fluorescein iodoacetamide. The affinity of these glycoclusters towards two plant lectins was assessed by surface plasmon resonance. The selectivity of their cell uptake was investigated by flow cytometry using two types of cells : a human hepatoma cell line (HepG2 cells) expressing the plasma membrane galactose-specific lectin, and monocyte-derived dendritic cells expressing the plasma membrane mannose-specific lectin. The glycoclusters containing four or five disaccharides were shown to bind plant lectins and cell surface membrane lectins with a narrow selectivity and with a high affinity.

In search of specific and highly selective sugar clusters for cell receptors, such as membrane lectins, various disaccharides were coupled to small peptide cores through an amide bond. In a first step, the reducing disaccharides, i.e. lactose and three different dimannoses, were converted into glycosyl-pyroglutamyl-ß-alanine derivatives. The free carboxylic group of these conjugates was then coupled to the a and epsilon amino groups of the core peptide (Lys(n)-Ala-Cys-NH2) with n = 1 to 5, with complete substitution leading to homogeneous glycoclusters. The thiol group of the cysteine residue was used to tag the glycosylated oligolysines upon reaction with fluorescein iodoacetamide. The affinity of these glycoclusters towards two plant lectins was assessed by surface plasmon resonance. The selectivity of their cell uptake was investigated by flow cytometry using two types of cells : a human hepatoma cell line (HepG2 cells) expressing the plasma membrane galactose-specific lectin, and monocyte-derived dendritic cells expressing the plasma membrane mannose-specific lectin. The glycoclusters containing four or five disaccharides were shown to bind plant lectins and cell surface membrane lectins with a narrow selectivity and with a high affinity.

Rondanino, C ; Bousser, MT ; Roche, AC ; Monsigny, M  (2002)

Nuclear import of glycoconjugates upon microinjection into the cytosol of living cells

Glycobiology 12 (10) 685-685 Art No. 128

Fajac, I ; Grosse, S ; Briand, P ; Monsigny, M  (2002)

Targeting of cell receptors and gene transfer efficiency : a balancing act

Gene Therapy 9 (11) 740-742
Vectors conjugated with ligands recognized by cell surface receptors are of interest for cystic fibrosis gene therapy since these vectors would allow cell-specific targeting. However, an efficient and specific uptake may be abrogated by a subsequent intracellular trafficking leading to an inefficient gene transfer. This has been shown for polylysine substituted with mannose residues. While mannose-specific membrane lectins are predominantly expressed at the surface of airway cells and mannosylated complexes are the most efficiently incorporated glycosylated complexes in these cells, mannosylated complexes lead to a low gene transfer efficiency because of an inefficient exit from endosomal compartments, a high accumulation in lysosomes and an inefficient nuclear import.

Vectors conjugated with ligands recognized by cell surface receptors are of interest for cystic fibrosis gene therapy since these vectors would allow cell-specific targeting. However, an efficient and specific uptake may be abrogated by a subsequent intracellular trafficking leading to an inefficient gene transfer. This has been shown for polylysine substituted with mannose residues. While mannose-specific membrane lectins are predominantly expressed at the surface of airway cells and mannosylated complexes are the most efficiently incorporated glycosylated complexes in these cells, mannosylated complexes lead to a low gene transfer efficiency because of an inefficient exit from endosomal compartments, a high accumulation in lysosomes and an inefficient nuclear import.

Kieda, C ; Paprocka, M ; Krawczenko, A ; Zalecki, P ; Dupuis, P ; Monsigny, M ; Radzikowski, C ; Dus, D  (2002)

New human microvascular endothelial cell lines with specific adhesion molecules phenotypes

Endothelium-New York 9 (4) 247-261
Vascular endothelial cells recognize blood-borne circulating cells and allow them to extravasate in a tissue-specific manner. Because this property determines the selectivity of lymphocyte homing, it is fundamental in physiological as well as pathological processes (inflammation, autoimmune diseases, metastasis). As a tool to assess the molecular basis of endothelium selectivity, microvascular endothelial cell lines of distinct tissue origin were established. Endothelial cells, isolated from lymphoid tissues (lymph nodes and appendix) and from nonlymphoid immune sites-intestine, lung, and skin-were immortalized in vitro. Their general endothelial characteristics, such as the presence of von Willebrand factor (wWf), angiotensin-converting enzyme (ACE), VE-cadherin, and the intracellular E-selectin, were preserved.

Vascular endothelial cells recognize blood-borne circulating cells and allow them to extravasate in a tissue-specific manner. Because this property determines the selectivity of lymphocyte homing, it is fundamental in physiological as well as pathological processes (inflammation, autoimmune diseases, metastasis). As a tool to assess the molecular basis of endothelium selectivity, microvascular endothelial cell lines of distinct tissue origin were established. Endothelial cells, isolated from lymphoid tissues (lymph nodes and appendix) and from nonlymphoid immune sites-intestine, lung, and skin-were immortalized in vitro. Their general endothelial characteristics, such as the presence of von Willebrand factor (wWf), angiotensin-converting enzyme (ACE), VE-cadherin, and the intracellular E-selectin, were preserved.


2001   Références trouvées : 6

Bedouet, L ; Bousser, MT ; Frison, N ; Boccaccio, C ; Abastado, JP ; Marceau, P ; Mayer, R ; Monsigny, M ; Roche, AC  (2001)

Uptake of dimannoside clusters and oligomannosides by human dendritic cells

Bioscience Reports 21 (6) 839-855
Knowing that human blood monocyte-derived dendritic cells express cell-surface mannose-specific lectins, we prepared various mannoses containing glycoconjugates with the aim of developing highly specific synthetic carriers of oligonucleotides and genes. Conjugates were prepared from oligosaccharides obtained by hydrazinolysis of Saccharomyces cerevisiae invertase glycopeptides.

Knowing that human blood monocyte-derived dendritic cells express cell-surface mannose-specific lectins, we prepared various mannoses containing glycoconjugates with the aim of developing highly specific synthetic carriers of oligonucleotides and genes. Conjugates were prepared from oligosaccharides obtained by hydrazinolysis of Saccharomyces cerevisiae invertase glycopeptides.

Meunier, L ; Monsigny, M ; Roche, AC  (2001)

Propynylated phosphodiester oligonucleotides inhibit ICAM-1 expression in A549 cells on electroporation

Antisense & Nucleic Acid Drug Development 11 (2) 117-123
Oligodeoxynucleotides (ODN) are used largely as either primers, antisense, or tripler-forming units. Phosphodiester ODN (PO-ODN), which are very rapidly degraded by exonucleases, must be protected at their ends. Even so, their life span inside cells is quite short. Phosphorothioate ODN (PS-ODN) are less sensitive to nucleases and are extensively used as antisense, Unfortunately, unlike PO-ODN, they interact with a number of molecules, including proteins, in addition to their specific nucleic acid targets, Their affinity for their target is lower than that of PO-ODN, PS-ODN containing propyne groups on C5 of pyrimidine have been shown to have a higher affinity toward their nucleic acid target. Here, we show that propynylated PO-ODN are more stable and much more efficient than their propyne-free counterparts. They are not efficient when they are used as lipoplexes, but they act as specific antisense on electroporation.

Oligodeoxynucleotides (ODN) are used largely as either primers, antisense, or tripler-forming units. Phosphodiester ODN (PO-ODN), which are very rapidly degraded by exonucleases, must be protected at their ends. Even so, their life span inside cells is quite short. Phosphorothioate ODN (PS-ODN) are less sensitive to nucleases and are extensively used as antisense, Unfortunately, unlike PO-ODN, they interact with a number of molecules, including proteins, in addition to their specific nucleic acid targets, Their affinity for their target is lower than that of PO-ODN, PS-ODN containing propyne groups on C5 of pyrimidine have been shown to have a higher affinity toward their nucleic acid target. Here, we show that propynylated PO-ODN are more stable and much more efficient than their propyne-free counterparts. They are not efficient when they are used as lipoplexes, but they act as specific antisense on electroporation.

Freulon, I ; Roche, AC ; Monsigny, M ; Mayer, R  (2001)

Delivery of oligonucleotides into mammalian cells by anionic peptides : comparison between monomeric and dimeric peptides

Biochemical Journal 354 671-679 Part 3
The use of antisense oligonucleotides as putative therapeutic agents is limited by their poor delivery into the cytosol and/or the nucleus because they are nut able to efficiently cross lipid bilayers. To circumvent this pitfall, anionic amphipathic peptides derived from the influenza virus fusogenic peptide have been used to destabilize membranes in an acidic environment. In this paper, we compare the ability of a monomeric and a dimeric peptide to introduce oligonucleotides into the cytosol and nuclei of several types of cultured cells. Cells incubated at pH 6.2 or at a slightly lower pH in the presence of the monomeric peptide but not the dimeric peptide were efficiently permeabilized. The location of fluorescent derivatives of peptides and of oligonucleotides was assessed by confocal microscopy. Both the peptides and oligonucleotides remained entrapped in vesicular compartments at neutral pH, at acidic pH, oligonucleotides in the presence of the monomeric peptide were mainly in the nucleus, while in the presence of the dimeric peptide they co-localized with the peptide into vesicles. The data are interpreted on the basis of the spectroscopic behaviour of monomeric and dimeric peptides in relation to the environmental pH.

The use of antisense oligonucleotides as putative therapeutic agents is limited by their poor delivery into the cytosol and/or the nucleus because they are nut able to efficiently cross lipid bilayers. To circumvent this pitfall, anionic amphipathic peptides derived from the influenza virus fusogenic peptide have been used to destabilize membranes in an acidic environment. In this paper, we compare the ability of a monomeric and a dimeric peptide to introduce oligonucleotides into the cytosol and nuclei of several types of cultured cells. Cells incubated at pH 6.2 or at a slightly lower pH in the presence of the monomeric peptide but not the dimeric peptide were efficiently permeabilized. The location of fluorescent derivatives of peptides and of oligonucleotides was assessed by confocal microscopy. Both the peptides and oligonucleotides remained entrapped in vesicular compartments at neutral pH, at acidic pH, oligonucleotides in the presence of the monomeric peptide were mainly in the nucleus, while in the presence of the dimeric peptide they co-localized with the peptide into vesicles. The data are interpreted on the basis of the spectroscopic behaviour of monomeric and dimeric peptides in relation to the environmental pH.

Roche, AC ; Monsigny, M ; Crocker, PR   (2001)

MR60/ERGIC-53, a mannose-specific shuttling intracellular membrane lectin

Results and Problems in Cell Differentiation. Mammalian Carbohydrate Recognition Systems 19-38

Monsigny, M  (2001)

Highlights : sugars make the decision. GlcNAcß-3Fuca-Thr(Ser) : a key motif in development.

Carbohydrate Letters 4 (2) 69

Fajac, I ; Briand, P ; Monsigny, M  (2001)

Gene therapy of cystic fibrosis : The glycofection approach

Glycoconjugate Journal 18 (9) 723-729
Many cells express surface membrane lectins that selectively bind and carry glycoconjugates into intracellular endosomes ; in addition, various intracellular membrane and soluble lectins act as shuttles between different compartments. On this basis, we developed glycosylated polycations, now called glycofectins (glycosylated polylysine and polyethyleneimine). Recently, we set up a simple way to transform oligosaccharides into glycosynthons suitable to substitute proteins or polymers. Glycofectins bind plasmid DNA leading to compact glycoplexes.

Many cells express surface membrane lectins that selectively bind and carry glycoconjugates into intracellular endosomes ; in addition, various intracellular membrane and soluble lectins act as shuttles between different compartments. On this basis, we developed glycosylated polycations, now called glycofectins (glycosylated polylysine and polyethyleneimine). Recently, we set up a simple way to transform oligosaccharides into glycosynthons suitable to substitute proteins or polymers. Glycofectins bind plasmid DNA leading to compact glycoplexes.


2000   Références trouvées : 7

Aubert, Y ; Bourgerie, S ; Meunier, L ; Mayer, R ; Roche, AC ; Monsigny, M ; Thuong, NT ; Asseline, U  (2000)

Optimized synthesis of phosphorothioate oligodeoxyribonucleotides substituted with a 5’-protected thiol function and a 3 ’-amino group

Nucleic Acids Research 28 (3) 818-825
A new deprotection procedure enables a medium scale preparation of phosphodiester and phosphorothioate oligonucleotides substituted with a protected thiol function at their 5'-ends and an amino group at their 3'-ends in good yield (up to 72 OD units/mu mol for a 19mer phosphorothioate). Syntheses of 3'-amino-substituted oligonucleotides were carried out on a modified support. A linker containing the thioacetyl moiety was manually coupled in two steps by first adding its phosphoramidite derivative in the presence of tetrazole followed by either oxidation or sulfurization to afford the bis-derivatized oligonucleotide bound to the support. Deprotection was achieved by treating the fully protected oligonucleotide with a mixture of 2,2'-dithiodipyridine and concentrated aqueous ammonia in the presence of phenol and methanol.

A new deprotection procedure enables a medium scale preparation of phosphodiester and phosphorothioate oligonucleotides substituted with a protected thiol function at their 5’-ends and an amino group at their 3’-ends in good yield (up to 72 OD units/mu mol for a 19mer phosphorothioate). Syntheses of 3’-amino-substituted oligonucleotides were carried out on a modified support. A linker containing the thioacetyl moiety was manually coupled in two steps by first adding its phosphoramidite derivative in the presence of tetrazole followed by either oxidation or sulfurization to afford the bis-derivatized oligonucleotide bound to the support. Deprotection was achieved by treating the fully protected oligonucleotide with a mixture of 2,2’-dithiodipyridine and concentrated aqueous ammonia in the presence of phenol and methanol.

Fajac, I ; Allo, JC ; Souil, E ; Merten, M ; Pichon, C ; Figarella, C ; Monsigny, M ; Briand, P ; Midoux, P  (2000)

Histidylated polylysine as a synthetic vector for gene transfer into immortalized cystic fibrosis airway surface and airway gland serous cells

Journal of Gene Medicine 2 (5) 368-378
Background : We recently designed a cationic polymer called histidylated polylysine made of polylysine partially substituted with histidyl residues which become protonated at slightly acidic pH. This polymer is thought to induce the leakage of acidic vesicles containing plasmid/histidylated polylysine complexes.

Background : We recently designed a cationic polymer called histidylated polylysine made of polylysine partially substituted with histidyl residues which become protonated at slightly acidic pH. This polymer is thought to induce the leakage of acidic vesicles containing plasmid/histidylated polylysine complexes.

Pichon, C ; Roufai, MB ; Monsigny, M ; Midoux, P  (2000)

Histidylated oligolysines increase the transmembrane passage and the biological activity of antisense oligonucleotides

Nucleic Acids Research 28 (2) 504-512
We have designed histidylated oligolysines which increase the uptake, the cytosolic delivery and the nuclear accumulation of antisense oligonucleotides (ODN), Flow cytometry analysis showed a 10-fold enhancement of the ODN uptake in the presence of histidylated oligolysines. The intracellular localizations of fluorescein-labeled ODN and of rhodamine-labeled histidylated oligolysines were investigated by confocal microscopy. Histidylated oligolysines favor the cytosolic delivery of ODN from endosomes and increase their nuclear accumulation.

We have designed histidylated oligolysines which increase the uptake, the cytosolic delivery and the nuclear accumulation of antisense oligonucleotides (ODN), Flow cytometry analysis showed a 10-fold enhancement of the ODN uptake in the presence of histidylated oligolysines. The intracellular localizations of fluorescein-labeled ODN and of rhodamine-labeled histidylated oligolysines were investigated by confocal microscopy. Histidylated oligolysines favor the cytosolic delivery of ODN from endosomes and increase their nuclear accumulation.

Monsigny, M ; Mayer, R ; Roche, A C  (2000)

Sugar-lectin interactions : sugar clusters, lectin multivalency and avidity.

Carbohydrate Letters 4 (1) 35-52

Freulon, I ; Monsigny, M ; Midoux, P ; Mayer, R  (2000)

Spacer length dependence on the efficiency of dimeric anionic peptides in gene transfer by glycosylated polylysine/plasmid complexes

Bioscience Reports 20 (5) 383-398
Amphiphilic anionic peptides have been used to enhance the efficiency of transfection by helping plasmids to escape from endosomes to the cytosol, It has been shown that efficiency of an eicosamers containing five glutamyl residues (E5), can be considerably enhanced either by transforming it into a dimer or by adding a tripeptide WYG in a C-terminal position (E5WYG). The dimerization of the peptide E5WYG leads to a more efficient tool when the dimerization device includes the tripeptide WYG unit and a longer spacer arm made of Gly-ß Ala-ß Ala residues. but to a 10-fold less efficient tool when the dimerization device includes a shorter spacer, a glycyl residue. Both dimers are taken up by the calls to a similar extent. Both dimers seem to be surrounded similarly as far as the environmental pH is concerned. In contrast, we found a correlation between the propensity of the peptides to adopt a helical structure at neutral pH and the gene transfer efficiency.

Amphiphilic anionic peptides have been used to enhance the efficiency of transfection by helping plasmids to escape from endosomes to the cytosol, It has been shown that efficiency of an eicosamers containing five glutamyl residues (E5), can be considerably enhanced either by transforming it into a dimer or by adding a tripeptide WYG in a C-terminal position (E5WYG). The dimerization of the peptide E5WYG leads to a more efficient tool when the dimerization device includes the tripeptide WYG unit and a longer spacer arm made of Gly-ß Ala-ß Ala residues. but to a 10-fold less efficient tool when the dimerization device includes a shorter spacer, a glycyl residue. Both dimers are taken up by the calls to a similar extent. Both dimers seem to be surrounded similarly as far as the environmental pH is concerned. In contrast, we found a correlation between the propensity of the peptides to adopt a helical structure at neutral pH and the gene transfer efficiency.

De Diesbach, P ; Berens, C ; N’Kuli, F ; Monsigny, M ; Sonveaux, E ; Wattiez, R ; Courtoy, PJ  (2000)

Identification, purification and partial characterisation of an oligonucleotide receptor in membranes of HepG2 cells

Nucleic Acids Research 28 (4) 868-874
The low and unpredictable uptake and cytosolic transfer of oligonucleotides (ODN) is a major reason for their limited benefit. Improving the ODN potential for therapy and research requires a better understanding of their receptor-mediated endocytosis. We have undertaken to identify a membrane ODN receptor on HepG2 cells by ligand blotting of cell extracts with [I-125]ODN and by photolabelling of living cells with a [I-125]ODN-benzophenone conjugate. A major band at 66 kDa was identified by the two methods.

The low and unpredictable uptake and cytosolic transfer of oligonucleotides (ODN) is a major reason for their limited benefit. Improving the ODN potential for therapy and research requires a better understanding of their receptor-mediated endocytosis. We have undertaken to identify a membrane ODN receptor on HepG2 cells by ligand blotting of cell extracts with [I-125]ODN and by photolabelling of living cells with a [I-125]ODN-benzophenone conjugate. A major band at 66 kDa was identified by the two methods.

Allo, JC ; Midoux, P ; Merten, M ; Souil, E ; Lipecka, J ; Figarella, C ; Monsigny, M ; Briand, P ; Fajac, I  (2000)

Efficient gene transfer into human normal and cystic fibrosis tracheal gland serous cells with synthetic vectors

American Journal of Respiratory Cell and Molecular Biology 22 (2) 166-175
Submucosal gland serous cells are believed to play a major role in the physiopathology of cystic fibrosis (CF) and may represent an important target for CF gene therapy. We have studied the efficiency of re porter gene transfer into immortalized normal (MM-39) and CF (CF-KM I) human airway epithelial gland serous cells using various synthetic vectors : glycosylated polylysines (glycofectins), polyethylenimine (PEI) (25 and 800 kD), lipofectin, and lipofectAMINE. In both cell lines, a high luciferase activity was achieved with various glycofectins, with PEI 25 kD, and with lipofectAMINE. After three transfections applied daily using a-glycosylated polylysine, 20% of the cells were transfected. At 24 h after CF trans membrane conductance regulator (CFTR) gene transfer into CF-KM4 cells using a-glycosylated poly lysine, the immunolocalization of CFTR was analyzed by laser scanning confocal microscopy and the transgenic CFTR was detected by an intense labeling of the plasma membrane.

Submucosal gland serous cells are believed to play a major role in the physiopathology of cystic fibrosis (CF) and may represent an important target for CF gene therapy. We have studied the efficiency of re porter gene transfer into immortalized normal (MM-39) and CF (CF-KM I) human airway epithelial gland serous cells using various synthetic vectors : glycosylated polylysines (glycofectins), polyethylenimine (PEI) (25 and 800 kD), lipofectin, and lipofectAMINE. In both cell lines, a high luciferase activity was achieved with various glycofectins, with PEI 25 kD, and with lipofectAMINE. After three transfections applied daily using a-glycosylated polylysine, 20% of the cells were transfected. At 24 h after CF trans membrane conductance regulator (CFTR) gene transfer into CF-KM4 cells using a-glycosylated poly lysine, the immunolocalization of CFTR was analyzed by laser scanning confocal microscopy and the transgenic CFTR was detected by an intense labeling of the plasma membrane.


1999   Références trouvées : 14

Carriere, V ; Piller, V ; Legrand, A ; Monsigny, M ; Roche, AC  (1999)

The sugar binding activity of MR60, a mannose-specific shuttling lectin, requires a dimeric state

Glycobiology 9 (10) 995-1002
MR60 is an intracellular membrane protein which has been shown to act as a mannoside specific lectin and to be identical to ERGIC-53, a protein characteristic of the endoplasmic reticulum-Golgi apparatus-intermediate compartment, acting as a shuttle. According to its primary sequence, this MR60/ERGIC-53 protein contains a luminal domain including the carbohydrate recognition domain, a stem, a transmembrane segment and a cytosolic domain. The endogenous MR60/ERGIC-53 protein is spontaneously oligomeric, (dimers and hexamers), In this paper, we study the relationship between the oligomerization state and the sugar binding capacity by using recombinant proteins.

MR60 is an intracellular membrane protein which has been shown to act as a mannoside specific lectin and to be identical to ERGIC-53, a protein characteristic of the endoplasmic reticulum-Golgi apparatus-intermediate compartment, acting as a shuttle. According to its primary sequence, this MR60/ERGIC-53 protein contains a luminal domain including the carbohydrate recognition domain, a stem, a transmembrane segment and a cytosolic domain. The endogenous MR60/ERGIC-53 protein is spontaneously oligomeric, (dimers and hexamers), In this paper, we study the relationship between the oligomerization state and the sugar binding capacity by using recombinant proteins.

Meunier, L ; Mayer, R ; Monsigny, M ; Roche, AC  (1999)

The nuclear export signal-dependent localization of oligonucleopeptides enhances the inhibition of the protein expression from a gene transcribed in cytosol

Nucleic Acids Research 27 (13) 2730-2736
Upon endocytosis, most oligodeoxynucleotides (ODNs) accumulate in vesicular compartments ; a tiny number of them cross the vesicle membrane, reach the cytosol and by passive diffusion enter the nucleus where they are entrapped. So far, the compartment in which an antisense ODN interacts with its mRNA target has not been precisely characterized. In an attempt to answer this question, ODN-peptides were designed with the aim of maintaining them in the cytosol, This has been achieved by a short peptide sequence called the nuclear export signal (NES). Upon microinjection, ODN-NES peptide conjugates were efficiently and rapidly exported from the nucleus to the cytosol whereas ODN-peptides containing an inactive NES were found to be located in the nucleus. The inhibitory activity of antisense ODN was tested in a system allowing the specific transcription of a luciferase reporter gene in the cytosol, Antisense propynylated ODN-NES peptide conjugates, directed against the luciferase gene, efficiently inhibited (75%) the cytosolic expression of luciferase whereas at the same concentration the peptide-free propynylated ODN or the propynylated ODN-peptides containing an inactive NES were nearly inactive.

Upon endocytosis, most oligodeoxynucleotides (ODNs) accumulate in vesicular compartments ; a tiny number of them cross the vesicle membrane, reach the cytosol and by passive diffusion enter the nucleus where they are entrapped. So far, the compartment in which an antisense ODN interacts with its mRNA target has not been precisely characterized. In an attempt to answer this question, ODN-peptides were designed with the aim of maintaining them in the cytosol, This has been achieved by a short peptide sequence called the nuclear export signal (NES). Upon microinjection, ODN-NES peptide conjugates were efficiently and rapidly exported from the nucleus to the cytosol whereas ODN-peptides containing an inactive NES were found to be located in the nucleus. The inhibitory activity of antisense ODN was tested in a system allowing the specific transcription of a luciferase reporter gene in the cytosol, Antisense propynylated ODN-NES peptide conjugates, directed against the luciferase gene, efficiently inhibited (75%) the cytosolic expression of luciferase whereas at the same concentration the peptide-free propynylated ODN or the propynylated ODN-peptides containing an inactive NES were nearly inactive.

Meunier, L ; Bourgerie, S ; Mayer, R ; Roche, AC ; Monsigny, M  (1999)

Optimized conditions to couple two water-soluble biomolecules through alkylamine thiolation and thioetherification

Bioconjugate Chemistry 10 (2) 206-212
A simple method far introducing, in buffered saline, a reactive sulfhydryl group on water-soluble molecules bearing an alkyl-amino group is described. This method is based on the use of two water-soluble reagents : 2-iminothiolane and 6,6'-dithiodinicotinic acid. The first one is open upon reaction with an amino group, and the generated thiol group is immediately protected by action of the second reagent. The optimal conditions were determined by taking into account the stability and the reactivity of both reagents with regards to pH and temperature. This method was validated through two applications, the substitution of bovine serum albumin with a bromoacetyl peptide and the substitution of an amino link at the 5' end of an oligonucleotide by reaction with either a fluorescent tag, iodoacetamidofluorescein, or a bromoacetyl peptide, upon reduction of the protected disulfide bridge with a third water-soluble reagent, namely tris(2-carboxyethyl)phosphine.

A simple method far introducing, in buffered saline, a reactive sulfhydryl group on water-soluble molecules bearing an alkyl-amino group is described. This method is based on the use of two water-soluble reagents : 2-iminothiolane and 6,6’-dithiodinicotinic acid. The first one is open upon reaction with an amino group, and the generated thiol group is immediately protected by action of the second reagent. The optimal conditions were determined by taking into account the stability and the reactivity of both reagents with regards to pH and temperature. This method was validated through two applications, the substitution of bovine serum albumin with a bromoacetyl peptide and the substitution of an amino link at the 5’ end of an oligonucleotide by reaction with either a fluorescent tag, iodoacetamidofluorescein, or a bromoacetyl peptide, upon reduction of the protected disulfide bridge with a third water-soluble reagent, namely tris(2-carboxyethyl)phosphine.

Carriere, V ; Landemarre, L ; Altemayer, V ; Motta, G ; Monsigny, M ; Roche, AC  (1999)

Intradermal DNA immunization : Antisera specific for the membrane lectin MR60/ERGIC-53

Bioscience Reports 19 (6) 559-569
The trafficking of intracellular membrane proteins in Golgi apparatus, endoplasmic reticulum or intermediate compartment has not yet been fully elucidated. The human MR60/ ERGIC-53 and the rat p58 proteins are one such protein ; and to study them in cell-free and in situ systems, high quality monospecific antisera are required. Highly specific antisera have been obtained after immunization of mice with plasmids containing a gene encoding either the full length or a truncated protein. The best results were obtained after intradermal injections of a plasmid encoding a truncated protein comprising both the luminal carbohydrate recognition domain and the stem down to a cysteine residue close to the C-terminal end, but neither the transmembrane nor the cytosolic domains. Such antisera have a very high titer and are very efficient tools to visualize the MR60 protein in situ or to selectively precipitate the MR60 proteins from a whole cell lysate.

The trafficking of intracellular membrane proteins in Golgi apparatus, endoplasmic reticulum or intermediate compartment has not yet been fully elucidated. The human MR60/ ERGIC-53 and the rat p58 proteins are one such protein ; and to study them in cell-free and in situ systems, high quality monospecific antisera are required. Highly specific antisera have been obtained after immunization of mice with plasmids containing a gene encoding either the full length or a truncated protein. The best results were obtained after intradermal injections of a plasmid encoding a truncated protein comprising both the luminal carbohydrate recognition domain and the stem down to a cysteine residue close to the C-terminal end, but neither the transmembrane nor the cytosolic domains. Such antisera have a very high titer and are very efficient tools to visualize the MR60 protein in situ or to selectively precipitate the MR60 proteins from a whole cell lysate.

Pichon, C ; Monsigny, M ; Roche, AC  (1999)

Intracellular localization of oligonucleotides : Influence of fixative protocols

Antisense & Nucleic Acid Drug Development 9 (1) 89-93
In many studies reporting the use of antisense oligonucleotides (ODN), the intracellular localization was investigated by using fluorescent-labeled oligonucleotides (F-ODN), More often, cells were fixed on uptake of F-ODN before microscopic analysis. We report here the influeuce of various methods of cell fixation on the intracellular localization of ODN, By confocal microscopy, we show that with unfixed cells, endocytosed peptides, oligonucleotides (M-r around 10,000), and endocytosed proteins were mainly localized in vesicular compartments. On mild fixation with paraformaldehyde, an identical intracellular localization was observed repeatedly after fixation, from immediately up to several days, In contrast, with methods based on the use of strong fixatives, such as methanol or acetone, the small molecules diffuse into the cytosol and in the case of oligonucleotides into the nucleus. These results point out the importance of the fixation protocol in the study of intracellular localization of ODN and their derivatives.

In many studies reporting the use of antisense oligonucleotides (ODN), the intracellular localization was investigated by using fluorescent-labeled oligonucleotides (F-ODN), More often, cells were fixed on uptake of F-ODN before microscopic analysis. We report here the influeuce of various methods of cell fixation on the intracellular localization of ODN, By confocal microscopy, we show that with unfixed cells, endocytosed peptides, oligonucleotides (M-r around 10,000), and endocytosed proteins were mainly localized in vesicular compartments. On mild fixation with paraformaldehyde, an identical intracellular localization was observed repeatedly after fixation, from immediately up to several days, In contrast, with methods based on the use of strong fixatives, such as methanol or acetone, the small molecules diffuse into the cytosol and in the case of oligonucleotides into the nucleus. These results point out the importance of the fixation protocol in the study of intracellular localization of ODN and their derivatives.

Monsigny, M ; Midoux, P ; Mayer, R ; Roche, AC  (1999)

Glycotargeting : Influence of the sugar moiety on both the uptake and the intracellular trafficking of nucleic acid carried by glycosylated polymers

Bioscience Reports 19 (2) 125-132
Nucleic acids (plasmids as well as oligonucleotides) used to specifically express or modulate the expression of a gene, must reach the cytosol and/or the nucleus. Several systems have been developed to increase their uptake and their efficiency. Glycosylated polylysines have been shown to specifically help nucleic acids to be taken up in cells expressing a given cell surface membrane lectin. However, it appeared that the efficiency of the imported nucleic acid was not directly related to the extent of the uptake. Indeed, some glycosylated polylysines bearing sugar moities which are poor ligands of the cell surface lectins of a given cell were found to be more efficient than those bearing better sugar ligands. The interpretation of this paradoxal result is discussed with regards to the nature of the compartment allowing the nucleic acid to cross the membrane and to be delivered in the cytosol on the one hand, and to the presence of intracellular lectins on the other hand.

Nucleic acids (plasmids as well as oligonucleotides) used to specifically express or modulate the expression of a gene, must reach the cytosol and/or the nucleus. Several systems have been developed to increase their uptake and their efficiency. Glycosylated polylysines have been shown to specifically help nucleic acids to be taken up in cells expressing a given cell surface membrane lectin. However, it appeared that the efficiency of the imported nucleic acid was not directly related to the extent of the uptake. Indeed, some glycosylated polylysines bearing sugar moities which are poor ligands of the cell surface lectins of a given cell were found to be more efficient than those bearing better sugar ligands. The interpretation of this paradoxal result is discussed with regards to the nature of the compartment allowing the nucleic acid to cross the membrane and to be delivered in the cytosol on the one hand, and to the presence of intracellular lectins on the other hand.

Fogel, S ; Guittaut, M ; Legrand, A ; Monsigny, M ; Hebert, E  (1999)

The Tat protein of HIV-1 induces galectin-3 expression

Glycobiology 9 (4) 383-387
Animal lectins play important roles in a variety of biological processes via their recognition of glycoconjugates. Galectin-3 is a ß-galactoside-binding lectin whose expression is associated with various pathological processes including human T lymphotropic virus (HTLV)-I-infection of human T cell lines and human immunodeficiency virus (HIV) infection of T-lymphoblastic Molt-3 cell line. In the case of HIV-infected cells, it has been suggested that the increase in galectin-3 expression could be related to the expression of the viral regulatory gene tat. These results prompt us to perform more extensive analyses of the relationship between galectin-3 and HIV-1 Tat expressions. In this study, we found that Tat protein expression induces an upregulation of galectin-3 in several human cell lines. In cotransfection experiments, the 5'-regulatory sequences of the galectin-3 gene were significantly upregulated by expression vectors encoding the Tat protein. Analysis performed with 5'-regulatory deleted sequences suggested that galectin-3 induction by Tat is dependent on activation of the Sp-1 binding transcription factor.

Animal lectins play important roles in a variety of biological processes via their recognition of glycoconjugates. Galectin-3 is a ß-galactoside-binding lectin whose expression is associated with various pathological processes including human T lymphotropic virus (HTLV)-I-infection of human T cell lines and human immunodeficiency virus (HIV) infection of T-lymphoblastic Molt-3 cell line. In the case of HIV-infected cells, it has been suggested that the increase in galectin-3 expression could be related to the expression of the viral regulatory gene tat. These results prompt us to perform more extensive analyses of the relationship between galectin-3 and HIV-1 Tat expressions. In this study, we found that Tat protein expression induces an upregulation of galectin-3 in several human cell lines. In cotransfection experiments, the 5’-regulatory sequences of the galectin-3 gene were significantly upregulated by expression vectors encoding the Tat protein. Analysis performed with 5’-regulatory deleted sequences suggested that galectin-3 induction by Tat is dependent on activation of the Sp-1 binding transcription factor.

Fajac, I ; Briand, P ; Monsigny, M ; Midoux, P  (1999)

Sugar-mediated uptake of glycosylated polylysines and gene transfer into normal and cystic fibrosis airway epithelial cells

Human Gene Therapy 10 (3) 395-406
We have examined the membrane lectin expressed by immortalized normal and cystic fibrosis (CF) airway epithelial cells, using fluorescein-labeled neoglycoproteins ; the uptake of plasmid DNA using fluoresceinylated glycoplexes (plasmid/glycosylated polylysine complexes) ; and the efficiency of gene transfer when glycosylated polylysines and glycosylated, partially gluconoylated polylysines were used as vectors, The most efficient up-take of neoglycoproteins by normal and CF cells was obtained with mannosylated BSA (bovine serum albumin). Similarly, the most efficient uptake of plasmid DNA was obtained with glycoplexes bearing a-D-Man residues.

We have examined the membrane lectin expressed by immortalized normal and cystic fibrosis (CF) airway epithelial cells, using fluorescein-labeled neoglycoproteins ; the uptake of plasmid DNA using fluoresceinylated glycoplexes (plasmid/glycosylated polylysine complexes) ; and the efficiency of gene transfer when glycosylated polylysines and glycosylated, partially gluconoylated polylysines were used as vectors, The most efficient up-take of neoglycoproteins by normal and CF cells was obtained with mannosylated BSA (bovine serum albumin). Similarly, the most efficient uptake of plasmid DNA was obtained with glycoplexes bearing a-D-Man residues.

Duverger, E ; Coppin, A ; Strecker, G ; Monsigny, M  (1999)

Interaction between lectins and neoglycoproteins containing new sialylated glycosynthons

Glycoconjugate Journal 16 (12) 793-800
Neoglycoconjugates are useful tools to study carbohydrate/protein interactions. In order to discover new lectins, to define their fine specificity or to study their intracellular trafficking, there is a need for neoglycoconjugates containing complex oligosaccharides. We recently set up a simple way to transform native oligosaccharides into glycosynthons. The present paper describes i) the synthesis of such glycosynthons starting with sialylated oligosides, ii) the preparation of sialylated neoglycoproteins and iii) their binding to sialic acid-specific lectins assessed by surface plasmon resonance experiments.

Neoglycoconjugates are useful tools to study carbohydrate/protein interactions. In order to discover new lectins, to define their fine specificity or to study their intracellular trafficking, there is a need for neoglycoconjugates containing complex oligosaccharides. We recently set up a simple way to transform native oligosaccharides into glycosynthons. The present paper describes i) the synthesis of such glycosynthons starting with sialylated oligosides, ii) the preparation of sialylated neoglycoproteins and iii) their binding to sialic acid-specific lectins assessed by surface plasmon resonance experiments.

Allo, JC ; Midoux, P ; Merten, M ; Monsigny, M ; Briand, P ; Figarella, C ; Fajac, I  (1999)

Histidylated polylysine/DNA complexes as efficient vectors for gene transfer into cystic fibrosis airway glandular serous cells.

American Journal of Respiratory and Critical Care Medicine 159 (3) A679-A679 Suppl. S

Kichler, A ; Freulon, I ; Boutin, V ; Mayer, R ; Monsigny, M ; Midoux, P  (1999)

Glycofection (TM) in the presence of anionic fusogenic peptides : A study of the parameters affecting the peptide-mediated enhancement of the transfection efficiency

Journal of Gene Medicine 1 (2) 134-143
Gene delivery mediated by polyplexes such as DNA complexed with polylysine conjugates is limited by the low efficiency of escape of DNA from the endosomes. One of the strategies which favors the transmembrane passage of polyplexes consists of adding anionic amphipathic peptides capable of destabilizing membranes in an acidic medium. Although less efficient than replication-defective adenoviruses, fusogenic peptides increase the expression of the reporter gene by a factor between 100 and 1000 depending on the cell line.

Gene delivery mediated by polyplexes such as DNA complexed with polylysine conjugates is limited by the low efficiency of escape of DNA from the endosomes. One of the strategies which favors the transmembrane passage of polyplexes consists of adding anionic amphipathic peptides capable of destabilizing membranes in an acidic medium. Although less efficient than replication-defective adenoviruses, fusogenic peptides increase the expression of the reporter gene by a factor between 100 and 1000 depending on the cell line.

Boutin, V ; Legrand, A ; Mayer, R ; Nachtigal, M ; Monsigny, M ; Midoux, P  (1999)

Glycofection : The ubiquitous roles of sugar bound on glycoplexes

Drug Delivery 6 (1) 45-50
Glycofection (transfection by using sugar-substituted polylysine) was assessed in order to provide an alternative to viral vectors for the transfer of genes into vascular smooth muscle cells, A rabbit vascular smooth muscle cell line (Rb-l cells) was selectively transfected by using glycoplexes (glycosylated polylysine/pSV2LUC complexes) in the presence of 10 mu M of the fusogenic peptide GALA, A sugar-specific transfection was obtained when the glycofection was conducted for 1 h with glycoplexes containing either a-Gal, a-Glc, a-GalNAc, ß-GlcNAc, or ß-GalNAc residues.

Glycofection (transfection by using sugar-substituted polylysine) was assessed in order to provide an alternative to viral vectors for the transfer of genes into vascular smooth muscle cells, A rabbit vascular smooth muscle cell line (Rb-l cells) was selectively transfected by using glycoplexes (glycosylated polylysine/pSV2LUC complexes) in the presence of 10 mu M of the fusogenic peptide GALA, A sugar-specific transfection was obtained when the glycofection was conducted for 1 h with glycoplexes containing either a-Gal, a-Glc, a-GalNAc, ß-GlcNAc, or ß-GalNAc residues.

Midoux, P ; Monsigny, M  (1999)

Efficient gene transfer by histidylated polylysine pDNA complexes

Bioconjugate Chemistry 10 (3) 406-411
Plasmid/polylysine complexes, which are used to transfect mammalian cells, increase the uptake of DNA, but plasmid molecules are sequestered into vesicles where they cannot escape to reach the nuclear machinery. However, the transfection efficiency increases when membrane-disrupting reagents such as chloroquine or fusogenic peptides, are used to disrupt endosomal membranes and to favor the delivery of plasmid into the cytosol. We designed a cationic polymer that forms complexes with a plasmid DNA (pDNA) and mediates the transfection of various cell lines in the absence of chloroquine or fusogenic peptides.

Plasmid/polylysine complexes, which are used to transfect mammalian cells, increase the uptake of DNA, but plasmid molecules are sequestered into vesicles where they cannot escape to reach the nuclear machinery. However, the transfection efficiency increases when membrane-disrupting reagents such as chloroquine or fusogenic peptides, are used to disrupt endosomal membranes and to favor the delivery of plasmid into the cytosol. We designed a cationic polymer that forms complexes with a plasmid DNA (pDNA) and mediates the transfection of various cell lines in the absence of chloroquine or fusogenic peptides.

Monsigny, M ; Pasternak, CA ; Surolia, A  (1999)

Bimal K Bachhawat (1925-1996) - Foreword

Bioscience Reports 19 (3) 155-156


1998   Références trouvées : 8

Quetard, C ; Bourgerie, S ; Normand-Sdiqui, N ; Mayer, R ; Strecker, G ; Midoux, P ; Roche, AC ; Monsigny, M  (1998)

Novel glycosynthons for glycoconjugate preparation : Oligosaccharylpyroglutamylanilide derivatives

Bioconjugate Chemistry 9 (2) 268-276
The reducing sugar of an oligosaccharide reacting with the a-amino group of an amino acid is converted to an N-oligosaccharylamino acid which can then be stabilized by N-acylation. Oligosaccharides in solution in N,N-dimethylformamide reacted with a-glutamyl-p-nitroanilide at 50 degrees C for a few hours, leading to an N-oligosaccharylglutamyl-p-nitroanilide. Then, the gamma-carboxylic group of the glutamyl moiety, activated by adding (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP), reacted with the substituted a-amino group of the glutamyl residue, leading to an N-oligosaccharylpyroglutamyl-p-nitroanilide within 0.5 h.

The reducing sugar of an oligosaccharide reacting with the a-amino group of an amino acid is converted to an N-oligosaccharylamino acid which can then be stabilized by N-acylation. Oligosaccharides in solution in N,N-dimethylformamide reacted with a-glutamyl-p-nitroanilide at 50 degrees C for a few hours, leading to an N-oligosaccharylglutamyl-p-nitroanilide. Then, the gamma-carboxylic group of the glutamyl moiety, activated by adding (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP), reacted with the substituted a-amino group of the glutamyl residue, leading to an N-oligosaccharylpyroglutamyl-p-nitroanilide within 0.5 h.

Monsigny, M ; Quetard, C ; Bourgerie, S ; Delay, D ; Pichon, C ; Midoux, P ; Mayer, R ; Roche, AC  (1998)

Glycotargeting : The preparation of glyco-amino acids and derivatives from unprotected reducing sugars

Biochimie 80 (2) 99-108
Lectins are present on the surface of many cells. Many lectins actively recycle from membrane to endosomes and efficiently take up glycoconjugates in a sugar-dependent manner. On this basis, glycoconjugates, specially those obtained by chemical means, are good candidates as carriers of drugs, oligonucleotides or genes. In this paper, we present a panel of methods suitable to transform unprotected reducing oligosaccharides into glycosynthons designed to be easily linked to therapeutic agents. All the glycosynthons presented here are glycosylamines or derivatives, mainly glyco-amino acids or glycopeptides. Glycosylamines are easy to obtain, but they are very labile in slightly acidic or neutral medium ; they must be stabilized, by acylation for instance. The coupling efficiency of a reducing sugar with ammonia as well as an alkylamine or an arylamine is higher at high temperature, however, because of the Amadori rearrangement, special conditions have to be selected to prepare the expected glycosylamine derivative with a high yield.

Lectins are present on the surface of many cells. Many lectins actively recycle from membrane to endosomes and efficiently take up glycoconjugates in a sugar-dependent manner. On this basis, glycoconjugates, specially those obtained by chemical means, are good candidates as carriers of drugs, oligonucleotides or genes. In this paper, we present a panel of methods suitable to transform unprotected reducing oligosaccharides into glycosynthons designed to be easily linked to therapeutic agents. All the glycosynthons presented here are glycosylamines or derivatives, mainly glyco-amino acids or glycopeptides. Glycosylamines are easy to obtain, but they are very labile in slightly acidic or neutral medium ; they must be stabilized, by acylation for instance. The coupling efficiency of a reducing sugar with ammonia as well as an alkylamine or an arylamine is higher at high temperature, however, because of the Amadori rearrangement, special conditions have to be selected to prepare the expected glycosylamine derivative with a high yield.

Kang, HC ; Ardourel, MY ; Guerin, B ; Monsigny, M ; Delmotte, FM  (1998)

Purification of two lectins from a nopalin Agrobacterium tumefaciens strain

Biochimie 80 (1) 87-94
Lectins were evidenced on the surface of one Agrobacterium tumefaciens wild strain (82.139) by agglutination test and neoglycoprotein labelling. Bacteria were incubated in the presence of various fluorescein-labelled neoglycoproteins and the binding was assessed by a fluorimetric method. Among the fluorescein-labelled neoglycoproteins tested, the one bearing a-D-galactosyl residues was the most efficient. The labelling was optimal at pH 5.0 and naught at pH above 7, The binding was specifically inhibited by homologous fluorescein-free neoglycoproteins. A galactoside-specific lectin was purified to homogeneity by affinity chromatography on agarose-A4 substituted with a-D-galactopyranosyl residues.

Lectins were evidenced on the surface of one Agrobacterium tumefaciens wild strain (82.139) by agglutination test and neoglycoprotein labelling. Bacteria were incubated in the presence of various fluorescein-labelled neoglycoproteins and the binding was assessed by a fluorimetric method. Among the fluorescein-labelled neoglycoproteins tested, the one bearing a-D-galactosyl residues was the most efficient. The labelling was optimal at pH 5.0 and naught at pH above 7, The binding was specifically inhibited by homologous fluorescein-free neoglycoproteins. A galactoside-specific lectin was purified to homogeneity by affinity chromatography on agarose-A4 substituted with a-D-galactopyranosyl residues.

Midoux, P ; Kichler, A ; Boutin, V ; Maurizot, JC ; Monsigny, M  (1998)

Membrane permeabilization and efficient gene transfer by a peptide containing several histidines

Bioconjugate Chemistry 9 (2) 260-267
We designed a peptide, H5WYG (GLFHAIAHFIHGGWHGLIHGWYG), that permeabilizes cell membrane at a slightly acidic pH but not at neutral pH. Absorbance, fluorescence, and circular dichroism spectra showed that H5WYG undergoes a dramatic conformational change between pH 7.0 and 6.0 that correlates with the protonation of the histidyl residues. Cell permeabilization studies monitored by flow cytometry on living cells showed that H5WYG permeabilizes the cell membrane with a great efficiency at pH 6.4 but was not active at neutral pH ; at pH 6.8, the peptide permeabilized 50% of the cells at 20 degrees C within 10 min. H5WYG increased the expression of genes transferred to cells as glycosylated polylysine-DNA complexes, and the transfection efficiency was not impaired in the presence of serum.

We designed a peptide, H5WYG (GLFHAIAHFIHGGWHGLIHGWYG), that permeabilizes cell membrane at a slightly acidic pH but not at neutral pH. Absorbance, fluorescence, and circular dichroism spectra showed that H5WYG undergoes a dramatic conformational change between pH 7.0 and 6.0 that correlates with the protonation of the histidyl residues. Cell permeabilization studies monitored by flow cytometry on living cells showed that H5WYG permeabilizes the cell membrane with a great efficiency at pH 6.4 but was not active at neutral pH ; at pH 6.8, the peptide permeabilized 50% of the cells at 20 degrees C within 10 min. H5WYG increased the expression of genes transferred to cells as glycosylated polylysine-DNA complexes, and the transfection efficiency was not impaired in the presence of serum.

Nachtigal, M ; Al-Assaad, Z ; Mayer, EP ; Kim, K ; Monsigny, M  (1998)

Galectin-3 expression in human atherosclerotic lesions

American Journal of Pathology 152 (5) 1199-1208
The expression of galectin-3, a ß-galactoside-binding lectin, was studied in atherosclerotic lesions from specimens obtained from carotid endarterectomies, lower limb amputations, and thoracic aortas from autopsies of young adult trauma victims, Immunohistochemical staining with the monoclonal antibody M3/38 demonstrated the presence of galectin-3 in advanced atherosclerotic lesions from each of 13 cases of carotid endarterectomy and 16 lower limb amputations and in the thoracic aorta of 4 of 20 cases of trauma victim adults, Immunostaining did not detect galectin-3 in umbilical cord and normal thoracic aorta arteries and limb veins.

The expression of galectin-3, a ß-galactoside-binding lectin, was studied in atherosclerotic lesions from specimens obtained from carotid endarterectomies, lower limb amputations, and thoracic aortas from autopsies of young adult trauma victims, Immunohistochemical staining with the monoclonal antibody M3/38 demonstrated the presence of galectin-3 in advanced atherosclerotic lesions from each of 13 cases of carotid endarterectomy and 16 lower limb amputations and in the thoracic aorta of 4 of 20 cases of trauma victim adults, Immunostaining did not detect galectin-3 in umbilical cord and normal thoracic aorta arteries and limb veins.

Nachtigal, M ; Al-Assaad, Z ; Mayer, EP ; Kim, K ; Movahed, R ; Monsigny, M  (1998)

Expression of galectin-3 in atherosclerotic vascular lesions.

Faseb Journal 12 (4) A482-A482 Part 1 Suppl. S

Bartholeyns, J ; Romet-Lemonne, JL ; Chokri, M ; Buyse, M ; Velu, T ; Bruyns, C ; Van de Winkel, JJ ; Heeney, J ; Koopman, G ; Malmsten, M ; De Groote, D ; Monsigny, M ; Midoux, P ; Alarcon, B  (1998)

Cellular vaccines

Research in Immunology 149 (7-8) 647-649
This project is devoted to the development of novel cellular vaccines designed to treat cancer patients. These cellular vaccines present and enhance immunogens, which will elicit a potent immune response. The goal is to achieve safe and effective immune reaction against the patient's own tumour. (1) Autologous cellular vaccines are prepared by processing circulating brood mononuclear cells outside of the patient's body (ex vivo) to differentiate them into antigen-presenting cells (APCs). Monocyte-derived APCs (MD-APCs) are then grown in the presence of exogenous target antigens (tumour cell debris, or apoptotic bodies) to become fully mature APCs. (2) Functionality for antigen presentation to T cells of ex vivo MD-APCs is evaluated in vivo. (3) Cellular vaccines are tested in selected rodent animal models. Efficiency and immune response are monitored in pertinent experimental systems for cancer. Pharmacological data are generated for clinical investigation.

This project is devoted to the development of novel cellular vaccines designed to treat cancer patients. These cellular vaccines present and enhance immunogens, which will elicit a potent immune response. The goal is to achieve safe and effective immune reaction against the patient’s own tumour. (1) Autologous cellular vaccines are prepared by processing circulating brood mononuclear cells outside of the patient’s body (ex vivo) to differentiate them into antigen-presenting cells (APCs). Monocyte-derived APCs (MD-APCs) are then grown in the presence of exogenous target antigens (tumour cell debris, or apoptotic bodies) to become fully mature APCs. (2) Functionality for antigen presentation to T cells of ex vivo MD-APCs is evaluated in vivo. (3) Cellular vaccines are tested in selected rodent animal models. Efficiency and immune response are monitored in pertinent experimental systems for cancer. Pharmacological data are generated for clinical investigation.

Florent, I ; Derhy, Z ; Allary, M ; Monsigny, M ; Mayer, R ; Schrevel, J  (1998)

A Plasmodium falciparum aminopeptidase gene belonging to the M1 family of zinc-metallopeptidases is expressed in erythrocytic stages

Molecular and Biochemical Parasitology 97 (1-2) 149-160
A new single copy gene has been isolated from Plasmodium falciparum, by immunoscreening a genomic DNA expression library. The gene appears devoid of introns, displays the classical A + T richness and codon usage of P. falciparum genes, and is transcribed into a 4 kb mRNA in erythrocytic stages. The deduced amino acid sequence corresponds to a 1056 residue protein (122 kDa) containing the canonical HExxHx(18)E signature of zinc-metallopeptidase active sites of the M1 family at position 467-490, a downstream conserved tyrosine residue involved in catalysis in position 551, and the GAMEN conserved motif characteristic of aminopeptidases in the M1 family, at position 431-435.

A new single copy gene has been isolated from Plasmodium falciparum, by immunoscreening a genomic DNA expression library. The gene appears devoid of introns, displays the classical A + T richness and codon usage of P. falciparum genes, and is transcribed into a 4 kb mRNA in erythrocytic stages. The deduced amino acid sequence corresponds to a 1056 residue protein (122 kDa) containing the canonical HExxHx(18)E signature of zinc-metallopeptidase active sites of the M1 family at position 467-490, a downstream conserved tyrosine residue involved in catalysis in position 551, and the GAMEN conserved motif characteristic of aminopeptidases in the M1 family, at position 431-435.


1997   Références trouvées : 13

Pichon, C ; Arar, K ; Stewart, AJ ; Dodon, MD ; Gazzolo, L ; Courtoy, PJ ; Mayer, R ; Monsigny, M ; Roche, AC  (1997)

Intracellular routing and inhibitory activity of oligonucleopeptides containing a KDEL motif

Molecular Pharmacology 51 (3) 431-438
On internalization, oligonucleotides (ODN) remain mostly sequestered in endocytic compartments. To increase their delivery into the cytosol and/or nucleus, which contain their targets, we attempted to guide them into compartments containing the KDEL receptor. Antisense ODN, phosphodiester protected at both ends, that are complementary to the AUG initiation site of gag(HIV-1) mRNA (odn) were linked to a peptide ending with the Lys-Asp-Glu-Leu (KDEL) motif in a carboxyl-terminal position (odn-p-KDEL) or with the Lys-Asp-Glu-Ala (odn-p-KDEA) as a control.

On internalization, oligonucleotides (ODN) remain mostly sequestered in endocytic compartments. To increase their delivery into the cytosol and/or nucleus, which contain their targets, we attempted to guide them into compartments containing the KDEL receptor. Antisense ODN, phosphodiester protected at both ends, that are complementary to the AUG initiation site of gag(HIV-1) mRNA (odn) were linked to a peptide ending with the Lys-Asp-Glu-Leu (KDEL) motif in a carboxyl-terminal position (odn-p-KDEL) or with the Lys-Asp-Glu-Ala (odn-p-KDEA) as a control.

Stewart, AJ ; Pichon, C ; Midoux, P ; Mayer, R ; Monsigny, M ; Roche, AC ; Aubertin, AM  (1997)

Fluorescent labelling of unmodified phosphorothioate oligodeoxynucleotides : Synthesis and characterization

New Journal of Chemistry 21 (1) 87-98
In this paper we describe the preparation and characterization of phosphorothioate oligodeoxynucleotides (pt-odn) substituted with a fluorescein molecule linked to the non-bridging sulfur of the internucleotidic linkage. This technique is original in that the starting material is a perphosphorothioate oligonucleotide. The phosphorothioate oligonucleotides, after reaction at pH 8.0 and 50 degrees C with iodoacetamido-fluorescein, yielded a fluorescent derivative, termed F-pt-odn. The two F-pt-odn used in this report contained 1.6 and 3.4 fluorescein per oligonucleotide and were found to be respectively 1.5 and 2.2 times more fluorescent than an alkylamidothiocarbamyl-fluoresceinyl-pt-odn (F-NH-pt-odn) containing a single reporter group.

In this paper we describe the preparation and characterization of phosphorothioate oligodeoxynucleotides (pt-odn) substituted with a fluorescein molecule linked to the non-bridging sulfur of the internucleotidic linkage. This technique is original in that the starting material is a perphosphorothioate oligonucleotide. The phosphorothioate oligonucleotides, after reaction at pH 8.0 and 50 degrees C with iodoacetamido-fluorescein, yielded a fluorescent derivative, termed F-pt-odn. The two F-pt-odn used in this report contained 1.6 and 3.4 fluorescein per oligonucleotide and were found to be respectively 1.5 and 2.2 times more fluorescent than an alkylamidothiocarbamyl-fluoresceinyl-pt-odn (F-NH-pt-odn) containing a single reporter group.

Pichon, C ; Freulon, I ; Midoux, P ; Mayer, R ; Monsigny, M ; Roche, AC  (1997)

Cytosolic and nuclear delivery of oligonucleotides mediated by an amphiphilic anionic peptide

Antisense & Nucleic Acid Drug Development 7 (4) 335-343
Antisense oligonucleotides (ODN) were easily introduced into the cytosol of mammalian cells on permeabilization of the plasma membrane by an amphiphilic anionic peptide. The E5CA peptide (GLFEAIAEFIEGG-WEGLIEGCA) is an E5 peptide analog derived from the N-terminal segment of the HA2 subunit of influenza virus hemagglutinin.

Antisense oligonucleotides (ODN) were easily introduced into the cytosol of mammalian cells on permeabilization of the plasma membrane by an amphiphilic anionic peptide. The E5CA peptide (GLFEAIAEFIEGG-WEGLIEGCA) is an E5 peptide analog derived from the N-terminal segment of the HA2 subunit of influenza virus hemagglutinin.

Condaminet, B ; Redziniak, G ; Monsigny, M ; Kieda, C  (1997)

Ultraviolet rays induced expression of lectins on the surface of a squamous carcinoma keratinocyte cell line

Experimental Cell Research 232 (2) 216-224
Human keratinocytic cells from squamous carcinoma (SCL-1) present, under resting conditions, relatively low amounts of endogenous lectins (sugar-binding proteins). Upon uv irradiation, they express on their cell surface large amounts of endogenous lectin molecules able to bind neoglycoproteins bearing either a-L-rhamnosyl or a-D-glucosyl residues, A similar binding specificity was found with normal human keratinocytes under the same culture conditions. At sunlike doses, uv.A (365 nm) was more efficient than uv.B (312 nm) in the expression of such receptors on the surface of SCL-1 cells, The increased presentation of lectins by SCL-1 cells was transient and reached a maximum 4 h after irradiation.

Human keratinocytic cells from squamous carcinoma (SCL-1) present, under resting conditions, relatively low amounts of endogenous lectins (sugar-binding proteins). Upon uv irradiation, they express on their cell surface large amounts of endogenous lectin molecules able to bind neoglycoproteins bearing either a-L-rhamnosyl or a-D-glucosyl residues, A similar binding specificity was found with normal human keratinocytes under the same culture conditions. At sunlike doses, uv.A (365 nm) was more efficient than uv.B (312 nm) in the expression of such receptors on the surface of SCL-1 cells, The increased presentation of lectins by SCL-1 cells was transient and reached a maximum 4 h after irradiation.

Erbacher, P ; Roche, AC ; Monsigny, M ; Midoux, P  (1997)

The reduction of the positive charges of polylysine by partial gluconoylation increases the transfection efficiency of polylysine/DNA complexes

Biochimica et Biophysica Acta-Biomembranes 1324 (1) 27-36
A polylysine partially substituted with polyhydroxyalkanoyl residues and specially with gluconoyl residues was developed in order to increase the transfection efficiency by decreasing the strength of the electrostatic interactions between the DNA and the cationic polymer. Partially gluconoylated polylysine/DNA complexes were more easily dissociated in solution and their transfection efficiency in the presence of chloroquine, evaluated with HepG2 cells, a human hepatocarcinoma line, was higher when 43 +/- 4% of the epsilon-amino groups of polylysine were blocked with gluconoyl residues.

A polylysine partially substituted with polyhydroxyalkanoyl residues and specially with gluconoyl residues was developed in order to increase the transfection efficiency by decreasing the strength of the electrostatic interactions between the DNA and the cationic polymer. Partially gluconoylated polylysine/DNA complexes were more easily dissociated in solution and their transfection efficiency in the presence of chloroquine, evaluated with HepG2 cells, a human hepatocarcinoma line, was higher when 43 +/- 4% of the epsilon-amino groups of polylysine were blocked with gluconoyl residues.

Hommel, M ; Attar, Z ; Fargeas, C ; Dourado, C ; Monsigny, M ; Mayer, R ; Chance, ML  (1997)

The direct agglutination test : a non-specific test specific for the diagnosis of visceral leishmaniasis ?

Annals of Tropical Medicine and Parasitology 91 (7) 795-802
Serology has an important role to play in the diagnosis of the severe clinical syndrome of visceral leishmaniasis (VL). The direct agglutination test (DAT), a simple agglutination test which requires no laboratory facilities, has become the preferred test, particularly for field studies. The nature of the antigens responsible for the agglutination of leishmanial promastigotes by the serum of VL patients is not known. A series of experiments which provide some clues to the molecular basis for the test and which indicate that there might be more in DAT than meets the eye is reported.

Serology has an important role to play in the diagnosis of the severe clinical syndrome of visceral leishmaniasis (VL). The direct agglutination test (DAT), a simple agglutination test which requires no laboratory facilities, has become the preferred test, particularly for field studies. The nature of the antigens responsible for the agglutination of leishmanial promastigotes by the serum of VL patients is not known. A series of experiments which provide some clues to the molecular basis for the test and which indicate that there might be more in DAT than meets the eye is reported.

Erbacher, P ; Roche, AC ; Monsigny, M ; Midoux, P  (1997)

Specific gene transfer based on biotinylated and gluconoylated polylysine/plasmid complexes

Drug Delivery 4 (3) 173-179
Complexes formed between plasmids and polylysine derivatives bearing recognition signals are the basis of nonviral vehicles suitable for gene delivery into eukaryotic cells by a receptor-mediated endocytosis process. We used an alternative procedure with the aim of achieving noncovalent attachment of recognition signals to plasmids. Biotinylated polylysine/DNA complexes were made between a plasmid and biotinylated polylysine by electrostatic interactions, and then the DNA complex was lactosylated via streptavidin bridges in order to target the galactose-specific membrane lectin of human hepatoma (HepG2) cells. HepG2 cells were efficiently transfected in a sugar-dependent manner with a polymer/DNA complex lactosylated with either biotinylated and lactosylated bovine serum albumin or lactosylated streptavidin.

Complexes formed between plasmids and polylysine derivatives bearing recognition signals are the basis of nonviral vehicles suitable for gene delivery into eukaryotic cells by a receptor-mediated endocytosis process. We used an alternative procedure with the aim of achieving noncovalent attachment of recognition signals to plasmids. Biotinylated polylysine/DNA complexes were made between a plasmid and biotinylated polylysine by electrostatic interactions, and then the DNA complex was lactosylated via streptavidin bridges in order to target the galactose-specific membrane lectin of human hepatoma (HepG2) cells. HepG2 cells were efficiently transfected in a sugar-dependent manner with a polymer/DNA complex lactosylated with either biotinylated and lactosylated bovine serum albumin or lactosylated streptavidin.

Gaudin, JC ; Arar, C ; Monsigny, M ; Legrand, A  (1997)

Modulation of the expression of the rabbit galectin-3 gene by p53 and c-Ha-ras proteins and PMA

Glycobiology 7 (8) 1089-1098
Galectin-3 is a galactose-binding lectin that has been found in several mammalian tissues, Galectin-3 gene is expressed in a wide range of normal and tumoral cells, In the case of myeloid cells, its expression correlates with the differentiation of monocytes to macrophages. In the case of cancer cell lines, its expression correlates with tumorigenicity and metastatic potential, The regulation of the expression of this gene is still largely unknown, The rabbit galectin-3 gene has been isolated and characterized, Its structure revealed an organization similar to that of the murine galectin-3 gene.

Galectin-3 is a galactose-binding lectin that has been found in several mammalian tissues, Galectin-3 gene is expressed in a wide range of normal and tumoral cells, In the case of myeloid cells, its expression correlates with the differentiation of monocytes to macrophages. In the case of cancer cell lines, its expression correlates with tumorigenicity and metastatic potential, The regulation of the expression of this gene is still largely unknown, The rabbit galectin-3 gene has been isolated and characterized, Its structure revealed an organization similar to that of the murine galectin-3 gene.

Raimond, J ; Zimonjic, DB ; Mignon, C ; Mattei, MG ; Popescu, NC ; Monsigny, M ; Legrand, A  (1997)

Mapping of the galectin-3 gene (LGALS3) to human Chromosome 14 at region 14q21-22

Mammalian Genome 8 (9) 706-707

Hebert, E ; Monsigny, M  (1997)

Lack of modulation of the galectin-3 and asialoglycoprotein receptor transcription in hepatocarcinoma of transgenic mice

Biology of The Cell 89 (7) 467-473
Galectin-3 and asialoglycoprotein receptor are lectins belonging to the classes of soluble lectins and of membrane C type lectins respectively. Conflicting results have been reported concerning their transcription level in the time course development of tumours. In the present study we investigated the abnormalities and the transcription levels of galectin-3 and asialoglycoprotein receptor genes in liver-targeted SV40 large T transgenic mice related to normal mice. in the strain expressing the highest level of large T, 100% of the male mice reproducibly developed an hepatocarcinoma.

Galectin-3 and asialoglycoprotein receptor are lectins belonging to the classes of soluble lectins and of membrane C type lectins respectively. Conflicting results have been reported concerning their transcription level in the time course development of tumours. In the present study we investigated the abnormalities and the transcription levels of galectin-3 and asialoglycoprotein receptor genes in liver-targeted SV40 large T transgenic mice related to normal mice. in the strain expressing the highest level of large T, 100% of the male mice reproducibly developed an hepatocarcinoma.

Pousset, D ; Piller, V ; Bureaud, N ; Monsigny, M ; Piller, F  (1997)

Increased a 2,6 sialylation of N-glycans in a transgenic mouse model of hepatocellular carcinoma

Cancer Research 57 (19) 4249-4256
Liver cancer is one of the most frequent and lethal malignancies worldwide, Early detection is hampered by the absence of reliable markers. Mice transgenic for the SV40 large T antigen under the control of a liver-specific promoter spontaneously develop well-differentiated hepatocellular carcinomas between 8 to 10 weeks of age. They are excellent models to investigate the alterations of protein expression in the early stages of tumor development and to follow these changes during tumor progression.

Liver cancer is one of the most frequent and lethal malignancies worldwide, Early detection is hampered by the absence of reliable markers. Mice transgenic for the SV40 large T antigen under the control of a liver-specific promoter spontaneously develop well-differentiated hepatocellular carcinomas between 8 to 10 weeks of age. They are excellent models to investigate the alterations of protein expression in the early stages of tumor development and to follow these changes during tumor progression.

Kollen, W ; Erbacher, P ; Midoux, P ; Roche, AC ; Monsigny, M ; Glick, MC ; Scanlin, TF  (1997)

Glycosylated polylysines - Nonviral vectors for gene transfer into cystic fibrosis airway epithelial cells

Chest 111 (6) S95-S96 Suppl. S

Stewart, AJ ; Pichon, C ; Meunier, L ; Midoux, P ; Monsigny, M ; Roche, AC  (1997)

Enhanced biological activity of antisense oligonucleotides complexed with glycosylated poly-L-lysine

Molecular Pharmacology 50 (6) 1487-1494
We sought to exploit glycosylated poly-L-lysine (pLK) to increase the uptake and biological antisense activity of a phosphorothioate oligonucleotide (pt-odn) [pt-odn complementary to the 3' noncoding region of intercellular adhesion molecule-1 ICAM-1)(odn(ICAM-1))] complementary to the 3'-noncoding region of ICAM-1 in A549 cells. Dose-dependent inhibition of ICAM-1 expression was obtained (IC50 = 500 nM) through treatment of cells with odn(ICAM-1) complexed with pLK carrying fucose residues in the presence of 100 mu M chloroquine. Alteration in the charge ratio between fucosylated pLK and pt-odn had a significant effect on the efficacy of inhibition (optimal conditions, charge ratio = 1.1).

We sought to exploit glycosylated poly-L-lysine (pLK) to increase the uptake and biological antisense activity of a phosphorothioate oligonucleotide (pt-odn) [pt-odn complementary to the 3’ noncoding region of intercellular adhesion molecule-1 ICAM-1)(odn(ICAM-1))] complementary to the 3’-noncoding region of ICAM-1 in A549 cells. Dose-dependent inhibition of ICAM-1 expression was obtained (IC50 = 500 nM) through treatment of cells with odn(ICAM-1) complexed with pLK carrying fucose residues in the presence of 100 mu M chloroquine. Alteration in the charge ratio between fucosylated pLK and pt-odn had a significant effect on the efficacy of inhibition (optimal conditions, charge ratio = 1.1).


1996   Références trouvées : 13

Hebert, E ; Roche, AC ; Nachtigal, M ; Monsigny, M  (1996)

Transformation but not ras-transfection increases the expression of galectin-3 in human HOS cells

Comptes Rendus de L Academie Des Sciences Serie Iii-Sciences de la Vie-Life Sciences 319 (10) 871-877
In ras-transfected NIH3T3 cells, the transcription of the Mr 34,000 ß-galactoside specific lectin galectin-3 depends on transformation phenotypes. This observation suggests that this lectin is associated with the transformation process and/or that the ras oncogene may modulate its expression ; nevertheless the involvement of ras-gene product in galectin-3 gene expression still remains unclear. In the present study, we investigated the galectin-3 expression in human HOS cells transiently or stably transfected with a ras-containing vector We observed an increase in galectin-3 mRNA and protein content in stably ras-transfected cells which had lost their anchorage dependence for growth but no increase in cells which needed anchorage for growth or in transiently ras-transfected cells These results suggest that the galectin-3 up-regulation in ras-transfected HOS cells is the consequence of the cell transformation rather than a divert effect of the ras gene product on galectin-3 gene expression.

In ras-transfected NIH3T3 cells, the transcription of the Mr 34,000 ß-galactoside specific lectin galectin-3 depends on transformation phenotypes. This observation suggests that this lectin is associated with the transformation process and/or that the ras oncogene may modulate its expression ; nevertheless the involvement of ras-gene product in galectin-3 gene expression still remains unclear. In the present study, we investigated the galectin-3 expression in human HOS cells transiently or stably transfected with a ras-containing vector We observed an increase in galectin-3 mRNA and protein content in stably ras-transfected cells which had lost their anchorage dependence for growth but no increase in cells which needed anchorage for growth or in transiently ras-transfected cells These results suggest that the galectin-3 up-regulation in ras-transfected HOS cells is the consequence of the cell transformation rather than a divert effect of the ras gene product on galectin-3 gene expression.

Erbacher, P ; Roche, AC ; Monsigny, M ; Midoux, P  (1996)

Putative role of chloroquine in gene transfer into a human hepatoma cell line by DNA lactosylated polylysine complexes

Experimental Cell Research 225 (1) 186-194
Chloroquine improves drastically the transfection of cells upon exposure to plasmid DNA/glycosylated polylysine complexes. So far the mechanism of action of chloroquine is not well understood. In this paper, the effect of chloroquine was investigated by measuring the transfection efficiency of a human hepatocarcinoma (HepG2 cells) by pSV2LUC/lactosylated polylysine complexes involving their internalization via the galactose-specific membrane lectin of these cells. The luciferase activity in the transfected cells was maximal when the transfection was performed for 3 or 4 h in the presence of 100 mu M chloroquine. The luciferase activity was also enhanced in the presence of primaquine, a chloroquine analogue, but was not increased when transfection was performed in the presence of ammonium chloride, methylamine, spermine, or monensin, compounds known to neutralize the pH of the endocytotic vesicle lumen as chloroquine does. Chloroquine enters cells and accumulates in vesicular compartments ; the overall intracellular concentration increases to 9 mM, which means that in the vesicular compartment, the chloroquine concentration is still higher. At such high concentrations, chloroquine induces the dissociation of plasmid DNA/lactosylated polylysine complexes, as shown in acellular experiments. (C) 1996 Academic Press, Inc.

Chloroquine improves drastically the transfection of cells upon exposure to plasmid DNA/glycosylated polylysine complexes. So far the mechanism of action of chloroquine is not well understood. In this paper, the effect of chloroquine was investigated by measuring the transfection efficiency of a human hepatocarcinoma (HepG2 cells) by pSV2LUC/lactosylated polylysine complexes involving their internalization via the galactose-specific membrane lectin of these cells. The luciferase activity in the transfected cells was maximal when the transfection was performed for 3 or 4 h in the presence of 100 mu M chloroquine. The luciferase activity was also enhanced in the presence of primaquine, a chloroquine analogue, but was not increased when transfection was performed in the presence of ammonium chloride, methylamine, spermine, or monensin, compounds known to neutralize the pH of the endocytotic vesicle lumen as chloroquine does. Chloroquine enters cells and accumulates in vesicular compartments ; the overall intracellular concentration increases to 9 mM, which means that in the vesicular compartment, the chloroquine concentration is still higher. At such high concentrations, chloroquine induces the dissociation of plasmid DNA/lactosylated polylysine complexes, as shown in acellular experiments. (C) 1996 Academic Press, Inc.

Kollen, WJW ; Erbacher, P ; Midoux, P ; Roche, AC ; Monsigny, M ; Glick, MC ; Scanlin, TF  (1996)

Nonviral vectors for gene transfer into cystic fibrosis (CF) airway epithelial cells.

Pediatric Research 39 (4) 2312-2312 Part 2

Duverger, E ; Roche, AC ; Monsigny, M  (1996)

N-acetylglucosamine-dependent nuclear import of neoglycoproteins

Glycobiology 6 (4) 381-386
Nuclear proteins bearing O-linked N-acetylglucosaminyl residues are involved in nuclear transport and in transcriptional processes, However, the role of glycosylation remains to be determined, It was proposed that glycosylation could be involved in macromolecular complex formation or in nuclear targeting, In the present study, we show that, in digitonin-permeabilized cells, BSA substituted with ß-di-N-acetylchitobioside (GlcNAc ß 4GlcNAc) is transported from the cytosol to the nucleus in a sugar dependent manner. The process is time and ATP dependent, Under the conditions used, the nuclear import of ß-di-N-acetyl-chitobios BSA, as it has also been previously shown for the nuclear import of a-glucosyl BSA (Duverger et al., J, Cell Sci., 108, 1325-1332, 1995) does not require the addition of a cytosolic extract, while the nuclear import of peptidic-NLS substituted BSA does require such an addition, These results suggest that GlcNAc can act as a nuclear localization signal.

Nuclear proteins bearing O-linked N-acetylglucosaminyl residues are involved in nuclear transport and in transcriptional processes, However, the role of glycosylation remains to be determined, It was proposed that glycosylation could be involved in macromolecular complex formation or in nuclear targeting, In the present study, we show that, in digitonin-permeabilized cells, BSA substituted with ß-di-N-acetylchitobioside (GlcNAc ß 4GlcNAc) is transported from the cytosol to the nucleus in a sugar dependent manner. The process is time and ATP dependent, Under the conditions used, the nuclear import of ß-di-N-acetyl-chitobios BSA, as it has also been previously shown for the nuclear import of a-glucosyl BSA (Duverger et al., J, Cell Sci., 108, 1325-1332, 1995) does not require the addition of a cytosolic extract, while the nuclear import of peptidic-NLS substituted BSA does require such an addition, These results suggest that GlcNAc can act as a nuclear localization signal.

Arar, C ; Mignon, C ; Mattei, MG ; Monsigny, M ; Roche, AC ; Legrand, A  (1996)

Mapping of the MR60/ERGIC-53 gene to human chromosome 18q21.3-18q22 by in situ hybridization

Mammalian Genome 7 (10) 791-792

Kollen, WJW ; Midoux, P ; Erbacher, P ; Yip, A ; Roche, AC ; Monsigny, M ; Glick, MC ; Scanlin, TF  (1996)

Gluconoylated and glycosylated polylysines as vectors for gene transfer into cystic fibrosis airway epithelial cells

Human Gene Therapy 7 (13) 1577-1586
To provide an alternative to viral vectors for the transfer of genes into airway epithelial cells in cystic fibrosis (CF), a novel set of substituted polylysines were employed, Polylysine was partially neutralized by blocking a number of positively charged residues with gluconoyl groups, In addition, polylysine was substituted with sugar residues on a specified number of amino groups, Using the gluconoylated polylysine as vector, the pCMVLuc plasmid gave high expression of the reporter gene luciferase in immortalized CF/T43 cells. The luciferase activity was 75-fold greater in the presence of 100 mu M chloroquine. Luciferase gene expression persisted at high levels for up to at least 120 hr following transfection.

To provide an alternative to viral vectors for the transfer of genes into airway epithelial cells in cystic fibrosis (CF), a novel set of substituted polylysines were employed, Polylysine was partially neutralized by blocking a number of positively charged residues with gluconoyl groups, In addition, polylysine was substituted with sugar residues on a specified number of amino groups, Using the gluconoylated polylysine as vector, the pCMVLuc plasmid gave high expression of the reporter gene luciferase in immortalized CF/T43 cells. The luciferase activity was 75-fold greater in the presence of 100 mu M chloroquine. Luciferase gene expression persisted at high levels for up to at least 120 hr following transfection.

Erbacher, P ; Bousser, MT ; Raimond, J ; Monsigny, M ; Midoux, P ; Roche, AC  (1996)

Gene transfer by DNA/glycosylated polylysine complexes into human blood monocyte-derived macrophages

Human Gene Therapy 7 (6) 721-729
Macrophages are putative target cells for expressing an exogenous gene with therapeutical effects. Knowing that macrophages express membrane lectins mediating endocytosis of their ligands, DNA/glycosylated polylysine complexes were used to transfect human blood monocyte-derived macrophages. Monocytes from human peripheral blood were matured in culture for 7 days to differentiate into macrophage-like cells in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF). Adherent cells, which displayed characteristic macrophage markers, CD 14, CD 11b, HLA-DR, and HLA-ABC antigens and mannose receptor, were transfected by DNA/glycosylated polylysine complexes in the presence of chloroquine.

Macrophages are putative target cells for expressing an exogenous gene with therapeutical effects. Knowing that macrophages express membrane lectins mediating endocytosis of their ligands, DNA/glycosylated polylysine complexes were used to transfect human blood monocyte-derived macrophages. Monocytes from human peripheral blood were matured in culture for 7 days to differentiate into macrophage-like cells in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF). Adherent cells, which displayed characteristic macrophage markers, CD 14, CD 11b, HLA-DR, and HLA-ABC antigens and mannose receptor, were transfected by DNA/glycosylated polylysine complexes in the presence of chloroquine.

Avrameas, A ; McIlroy, D ; Hosmalin, A ; Autran, B ; Debre, P ; Monsigny, M ; Roche, AC ; Midoux, P  (1996)

Expression of a mannose/fucose membrane lectin on human dendritic cells

European Journal of Immunology 26 (2) 394-400
Dendritic cells (DC) are the most efficient antigen presenting cells for T lymphocytes. CD1a CD14(-) DC with high antigen-presenting capacities can now be obtained easily from adherent peripheral blood monocytes by culture in the presence of granulocyte/macrophage colony-stimulating factor and interleukin-4 (Sallusto et al.. J. Exp. Med. 1994, 1795, 1109). Human macrophages express a membrane lectin. or sugar-specific receptor. which specifically mediates the binding and endocytosis of mannose- and fucose-terminated glycoproteins and is involved in the phagocytosis of pathogens.

Dendritic cells (DC) are the most efficient antigen presenting cells for T lymphocytes. CD1a CD14(-) DC with high antigen-presenting capacities can now be obtained easily from adherent peripheral blood monocytes by culture in the presence of granulocyte/macrophage colony-stimulating factor and interleukin-4 (Sallusto et al.. J. Exp. Med. 1994, 1795, 1109). Human macrophages express a membrane lectin. or sugar-specific receptor. which specifically mediates the binding and endocytosis of mannose- and fucose-terminated glycoproteins and is involved in the phagocytosis of pathogens.

Itin, C ; Roche, AC ; Monsigny, M ; Hauri, HP  (1996)

ERGIC-53 is a functional mannose-selective and calcium-dependent human homologue of leguminous lectins

Molecular Biology of The Cell 7 (3) 483-493
Based on sequence homologies with leguminous lectins, the intermediate compartment marker ERGIC-53 was proposed to be a member of a putative new class of animal lectins associated with the secretory pathway. Independently, a promyelocytic protein, MR60, was purified by mannose-column chromatography, and a cDNA was isolated that matched MR60 peptide sequences. This cDNA was identical to that of ERGIC-53 and homologies With the animal lectin family of the galectins were noticed. Not all peptide sequences of MR60, however, were found in ERGIC-53, raising the possibility that another protein associated with ERGIC-53 may possess the lectin activity. Here, we provide the first direct evidence for a lectin function of ERGIC-53.

Based on sequence homologies with leguminous lectins, the intermediate compartment marker ERGIC-53 was proposed to be a member of a putative new class of animal lectins associated with the secretory pathway. Independently, a promyelocytic protein, MR60, was purified by mannose-column chromatography, and a cDNA was isolated that matched MR60 peptide sequences. This cDNA was identical to that of ERGIC-53 and homologies With the animal lectin family of the galectins were noticed. Not all peptide sequences of MR60, however, were found in ERGIC-53, raising the possibility that another protein associated with ERGIC-53 may possess the lectin activity. Here, we provide the first direct evidence for a lectin function of ERGIC-53.

Fargeas, C ; Hommel, M ; Maingon, R ; Dourado, C ; Monsigny, M ; Mayer, R  (1996)

Synthetic peptide-based enzyme-linked immunosorbent assay for serodiagnosis of visceral leishmaniasis

Journal of Clinical Microbiology 34 (2) 241-248
Synthetic peptides, derived from the amino acid sequence of a Leishmania donovani clone, were used to develop an enzyme-linked immunosorbent assay (ELISA) for detecting antibodies against L. donovani. For this purpose, five peptides were conjugated to a protein carrier, human serum albumin (HSA), by using a heterobifunctional reagent, epsilon-maleimidocaproic acid N-hydrolrysuccinimide ester, to obtain a well-defined product. The sensitivity and the specificity of the peptide-specific ELISA were determined with a panel of 106 serum samples from individuals living in areas where visceral leishmaniasis is endemic ; sera from post-kala azar dermal leishmaniasis-infected patients and from individuals suffering from other infectious diseases were also included.

Synthetic peptides, derived from the amino acid sequence of a Leishmania donovani clone, were used to develop an enzyme-linked immunosorbent assay (ELISA) for detecting antibodies against L. donovani. For this purpose, five peptides were conjugated to a protein carrier, human serum albumin (HSA), by using a heterobifunctional reagent, epsilon-maleimidocaproic acid N-hydrolrysuccinimide ester, to obtain a well-defined product. The sensitivity and the specificity of the peptide-specific ELISA were determined with a panel of 106 serum samples from individuals living in areas where visceral leishmaniasis is endemic ; sera from post-kala azar dermal leishmaniasis-infected patients and from individuals suffering from other infectious diseases were also included.

Monsigny, Michel ; Midoux, Patrick ; Roche, Annie-Claude ; Legrand, Alain ; Mayer, Roger  (1996)

Selective transfection of animal cells by non viral vectors, polylysine carrying recognition signals

Comptes Rendus Des Seances de la Societe de Biologie et de Ses Filiales 190 (1) 39-43
A process for the selective transfer of genes using glycosylated polylysine was established. Glycosylated polylysines and plasmid DNA form complexes which are taken up by cells expressing surface lectins recognizing the sugar moieties of glycosylated polylysines. Modifications made to this process allow for efficient and specific transfer in in vitro animal cell models.

A process for the selective transfer of genes using glycosylated polylysine was established. Glycosylated polylysines and plasmid DNA form complexes which are taken up by cells expressing surface lectins recognizing the sugar moieties of glycosylated polylysines. Modifications made to this process allow for efficient and specific transfer in in vitro animal cell models.

Denis, V ; Dupuis, P ; Bizouarne, N ; Sampaio, SD ; Hong, L ; Lebret, M ; Monsigny, M ; Nakache, M ; Kieda, C  (1996)

Selective induction of peripheral and mucosal endothelial cell addressins with peripheral lymph nodes and Peyer’s patch cell-conditioned media

Journal of Leukocyte Biology 60 (6) 744-752
Vascular endothelial cell addressins play an important role in lymphocyte homing in secondary lymphoid organs and in chronic inflammatory areas, A SV40 large T antigen-immortalized cell line from peripheral lymph nodes, HECa10 [Bizouarne et al., 1993a], was used to characterize the location of addressins with regard to environmental factors and cytokines. For this purpose, two monoclonal antibodies, MECA 79 and MECA 367, specific for peripheral lymph node vascular addressin and for mucosal addressin (Peyer's patches), respectively, were bound to unstimulated HECa10 cells. Both mucosal and peripheral addressins were detected inside the cells and in cellular extracts of the resting cells. On the cell surface, both addressins could be evidenced on the same cells at a moderate level of expression.

Vascular endothelial cell addressins play an important role in lymphocyte homing in secondary lymphoid organs and in chronic inflammatory areas, A SV40 large T antigen-immortalized cell line from peripheral lymph nodes, HECa10 [Bizouarne et al., 1993a], was used to characterize the location of addressins with regard to environmental factors and cytokines. For this purpose, two monoclonal antibodies, MECA 79 and MECA 367, specific for peripheral lymph node vascular addressin and for mucosal addressin (Peyer’s patches), respectively, were bound to unstimulated HECa10 cells. Both mucosal and peripheral addressins were detected inside the cells and in cellular extracts of the resting cells. On the cell surface, both addressins could be evidenced on the same cells at a moderate level of expression.

Watier, H ; Guillaumin, JM ; Piller, F ; Lacord, M ; Thibault, G ; Lebranchu, Y ; Monsigny, M ; Bardos, P  (1996)

Removal of terminal a-galactosyl residues from xenogeneic porcine endothelial cells - Decrease in complement-mediated cytotoxicity but persistence of IgG1-mediated antibody-dependent cell-mediated cytotoxicity

Transplantation 62 (1) 105-113
To determine the role of the terminal a-galactosyl residue in the endothelial damage mediated by human xenoreactive natural antibodies (IgM and IgG), we treated porcine endothelial cells in culture with green coffee bean a-galactosidase. A practically complete removal of terminal a-Gal residues (as evaluated by flow cytometry with Bandeiraea simplicifolia isolectin B4) and concomitant exposure of N-acetyllactosamine were obtained without altering cell viability. A dramatic decrease in IgM and IgG binding (from a pool of human sera) was observed, confirming the key role of the a-galactosyl residues. The enzyme treatment did not induce any nonspecific immunoglobulin binding sites, but led to the exposure of new epitopes for a minor fraction of IgM. The main residual IgM and IgG binding could be due to xenoantigens other than the a-galactosyl residues.

To determine the role of the terminal a-galactosyl residue in the endothelial damage mediated by human xenoreactive natural antibodies (IgM and IgG), we treated porcine endothelial cells in culture with green coffee bean a-galactosidase. A practically complete removal of terminal a-Gal residues (as evaluated by flow cytometry with Bandeiraea simplicifolia isolectin B4) and concomitant exposure of N-acetyllactosamine were obtained without altering cell viability. A dramatic decrease in IgM and IgG binding (from a pool of human sera) was observed, confirming the key role of the a-galactosyl residues. The enzyme treatment did not induce any nonspecific immunoglobulin binding sites, but led to the exposure of new epitopes for a minor fraction of IgM. The main residual IgM and IgG binding could be due to xenoantigens other than the a-galactosyl residues.


1995   Références trouvées : 11

Sdiqui, N., Arar, K., Midoux, P., Monsigny, M. & Ropche, A.C.  (1995)

Inhibition of human mammary cell line proliferation by membrane lectin-mediated uptake of Ha-ras antisense oligonucleotide

Drug Delivery 2, 63-72

Arar, K ; Aubertin, AM ; Roche, AC ; Monsigny, M ; Mayer, R  (1995)

Synthesis and antiviral activity of peptide-oligonucleotide conjugates prepared by using n-a-(bromoacetyl)peptides

Bioconjugate Chemistry 6 (5) 573-577
Antisense oligonucleotides represent an interesting tool for selective inhibition of gene expression. In order to direct oligonucleotides to specific compartments within the cell, we have investigated the possibility of coupling them to a signal peptide Lys-Asp-Glu-Leu (KDEL). This sequence should be able to convey oligonucleotides to the endoplasmic reticulum and from there to the cytosol and the nucleus where their targets are located. On this basis we prepared peptide-oligonucleotide conjugates by coupling, in a single step, a N-a-bromoacetyl peptide with an oligonucleotide bearing a thiol group, through a thioether bond. This paper deals with the definition of the optimal pH and temperature conditions leading to an efficient synthesis of peptide-oligonucleotide conjugates : the reaction was quantitative at pH 7.5 within few hours.

Antisense oligonucleotides represent an interesting tool for selective inhibition of gene expression. In order to direct oligonucleotides to specific compartments within the cell, we have investigated the possibility of coupling them to a signal peptide Lys-Asp-Glu-Leu (KDEL). This sequence should be able to convey oligonucleotides to the endoplasmic reticulum and from there to the cytosol and the nucleus where their targets are located. On this basis we prepared peptide-oligonucleotide conjugates by coupling, in a single step, a N-a-bromoacetyl peptide with an oligonucleotide bearing a thiol group, through a thioether bond. This paper deals with the definition of the optimal pH and temperature conditions leading to an efficient synthesis of peptide-oligonucleotide conjugates : the reaction was quantitative at pH 7.5 within few hours.

Duverger, E ; Pellerinmendes, C ; Mayer, R ; Roche, AC ; Monsigny, M  (1995)

Nuclear import of glycoconjugates is distinct from the classical nls pathway

Journal of Cell Science 108 1325-1332 Part 4
The nuclear import of many proteins depends on a short peptide sequence called the nuclear localization signal, However, glycosylated proteins, which lack such a nuclear localization signal, upon their injection into the cytosol by electroporation, enter the nucleus in a sugar-dependent manner. This paper brings new insights on the mechanism of this process, based on a study of neoglycoprotein nuclear uptake by digitonin-permeabilized cells. The nuclear import of neoglycoproteins is energy dependent : it does not occur when cells are maintained at 4 degrees C or when cells are ATP-depleted by treatment with apyrase.

The nuclear import of many proteins depends on a short peptide sequence called the nuclear localization signal, However, glycosylated proteins, which lack such a nuclear localization signal, upon their injection into the cytosol by electroporation, enter the nucleus in a sugar-dependent manner. This paper brings new insights on the mechanism of this process, based on a study of neoglycoprotein nuclear uptake by digitonin-permeabilized cells. The nuclear import of neoglycoproteins is energy dependent : it does not occur when cells are maintained at 4 degrees C or when cells are ATP-depleted by treatment with apyrase.

Santambien, P ; Sdiqui, S ; Hubert, E ; Girot, P ; Roche, AC ; Monsigny, M ; Boschetti, E  (1995)

In vitro toxicity assays for dye ligands used in affinity-chromatography

Journal of Chromatography B-Biomedical Applications 664 (1) 241-246
Some reactive textile dyes have been used for years as biomimetic ligands in protein purification. There has been reluctance, however, to use these dyes on a large scale for therapeutically applicable proteins for fear of possible dye leakage and consequent contamination. Therefore, toxicological data are necessary to quantify the level of this hazard. This study deals with a series of in vitro toxicity investigations with eukaryotic cells (growth, polyploidy, etc.) and with prokaryotic cells (Escherichia coli) for genotoxic studies. Both approaches demonstrated a lack of or slight toxicity for Reactive Blue 2 and Reactive Red 120 and their derivatives over the range 10-62.5 mu g/ml in several assays.

Some reactive textile dyes have been used for years as biomimetic ligands in protein purification. There has been reluctance, however, to use these dyes on a large scale for therapeutically applicable proteins for fear of possible dye leakage and consequent contamination. Therefore, toxicological data are necessary to quantify the level of this hazard. This study deals with a series of in vitro toxicity investigations with eukaryotic cells (growth, polyploidy, etc.) and with prokaryotic cells (Escherichia coli) for genotoxic studies. Both approaches demonstrated a lack of or slight toxicity for Reactive Blue 2 and Reactive Red 120 and their derivatives over the range 10-62.5 mu g/ml in several assays.

Erbacher, P ; Roche, AC ; Monsigny, M ; Midoux, P  (1995)

Glycosylated polylysine DNA complexes - gene-transfer efficiency in relation with the size and the sugar substitution level of glycosylated polylysines and with the plasmid size

Bioconjugate Chemistry 6 (4) 401-410
A DNA delivery system based on the use of polylysine substituted with small recognition signals, such as carbohydrate moieties specifically recognized by membrane lectins present in a given cell line, has been developed [Midoux et al. (1993) Nucleic Acids Res. 21, 871-878]. Human hepatoma (HepG2) cells which express a galactose-specific membrane lectin are efficiently transfected in the presence of chloroquine with pSV2Luc plasmid complexed with a lactosylated polylysine.

A DNA delivery system based on the use of polylysine substituted with small recognition signals, such as carbohydrate moieties specifically recognized by membrane lectins present in a given cell line, has been developed [Midoux et al. (1993) Nucleic Acids Res. 21, 871-878]. Human hepatoma (HepG2) cells which express a galactose-specific membrane lectin are efficiently transfected in the presence of chloroquine with pSV2Luc plasmid complexed with a lactosylated polylysine.

Arar, C ; Carpentier, V ; Lecaer, JP ; Monsigny, M ; Legrand, A ; Roche, AC  (1995)

Ergic-53, a membrane-protein of the endoplasmic reticulum-golgi intermediate compartment, is identical to mr60, an intracellular mannose-specific lectin of myelomonocytic cells

Journal of Biological Chemistry 270 (8) 3551-3553
A mannose specific membrane lectin (MR60) isolated from human myelomonocytic HL60 cells by affinity chromatography is expressed in intracellular organelles of immature monocytes (Pimpaneau, V., Midoux, P., Monsigny, M., and Roche, A. C. (1991) Carbohydr. Res. 213, 95-108). It is not present at the cell surface and is immunochemically and structurally distinct from the M(r) 175,000 mannose receptor of mature macrophages. MR60 cDNA was isolated and characterized ; on the basis of its sequence, MR60 is not related to any known mammalian lectins.

A mannose specific membrane lectin (MR60) isolated from human myelomonocytic HL60 cells by affinity chromatography is expressed in intracellular organelles of immature monocytes (Pimpaneau, V., Midoux, P., Monsigny, M., and Roche, A. C. (1991) Carbohydr. Res. 213, 95-108). It is not present at the cell surface and is immunochemically and structurally distinct from the M(r) 175,000 mannose receptor of mature macrophages. MR60 cDNA was isolated and characterized ; on the basis of its sequence, MR60 is not related to any known mammalian lectins.

Rouleux-Bonnin, F ; Monsigny, M ; Legrand, A  (1995)

Transcriptional expression of mannose receptor gene during differentiation of human macrophages

Biochemical and Biophysical Research Communications 217 (1) 106-112
Mannose receptor is a differentiation marker of macrophages. Circulating monocytes isolated from plasma are devoid of this receptor ; upon culture this receptor is rapidly expressed. Its expression is modulated by a variety of inflammatory and antiinflammatory agents. In the present study, we investigated its transcription level during the differentiation process. Mannose receptor mRNA was monitored by quantitative RT-PCR on freshly harvested monocytes and on monocytes cultivated up to four days. No transcription was detected in freshly harvested cells, the transcription increased during the first 24 h upon adhesion and then decreased. (C) 1995 Academic Press, Inc.

Mannose receptor is a differentiation marker of macrophages. Circulating monocytes isolated from plasma are devoid of this receptor ; upon culture this receptor is rapidly expressed. Its expression is modulated by a variety of inflammatory and antiinflammatory agents. In the present study, we investigated its transcription level during the differentiation process. Mannose receptor mRNA was monitored by quantitative RT-PCR on freshly harvested monocytes and on monocytes cultivated up to four days. No transcription was detected in freshly harvested cells, the transcription increased during the first 24 h upon adhesion and then decreased. (C) 1995 Academic Press, Inc.

Raimond, J ; Rouleux, F ; Monsigny, M ; Legrand, A  (1995)

The 2nd intron of the human galectin-3 gene has a strong promoter activity down-regulated by P53

Febs Letters 363 (1-2) 165-169
Galectin-3 is a galactose-specific lectin which has been shown to be involved in several biological functions such as cell growth regulation, cell aggregation and cell differentiation. The partial cloning of the human genomic sequences reveals the presence of a 651 bp intron, 18 bp downstream of the translation initiation site, This intron contains several regulatory elements found in many eukaryotic genes, This sequence, when inserted upstream of a promoter-free luciferase gene, induces the expression of luciferase, demonstrating the promoter activity of the intron upon transfection in human or murine cells, This promoter activity is down-modulated by wild-type p53 but not by a mutated form of p53.

Galectin-3 is a galactose-specific lectin which has been shown to be involved in several biological functions such as cell growth regulation, cell aggregation and cell differentiation. The partial cloning of the human genomic sequences reveals the presence of a 651 bp intron, 18 bp downstream of the translation initiation site, This intron contains several regulatory elements found in many eukaryotic genes, This sequence, when inserted upstream of a promoter-free luciferase gene, induces the expression of luciferase, demonstrating the promoter activity of the intron upon transfection in human or murine cells, This promoter activity is down-modulated by wild-type p53 but not by a mutated form of p53.

Midoux, P ; Mayer, R ; Monsigny, M  (1995)

Membrane permeabilization by a-helical peptides - a flow-cytometry study

Biochimica et Biophysica Acta-Biomembranes 1239 (2) 249-256
The permeabilization by a-helical peptides of nucleated mammalian cells can be monitored by flow cytometry. Ethidium bromide, a non fluorescent and poorly membrane permeant molecule, becomes strongly fluorescent only upon binding to DNA. On this basis, the permeabilization of the plasma membrane of HL60 promyelocytic cells induced by a-helical peptides such as melittin, succinylated melittin and anionic peptides derived from the N-terminus of HA2 subunit of the influenza virus hemagglutinin, was measured. Melittin (GIGAVLKVLTTGLPALISWIKRKRQQ-NH2) caused a rapid (

The permeabilization by a-helical peptides of nucleated mammalian cells can be monitored by flow cytometry. Ethidium bromide, a non fluorescent and poorly membrane permeant molecule, becomes strongly fluorescent only upon binding to DNA. On this basis, the permeabilization of the plasma membrane of HL60 promyelocytic cells induced by a-helical peptides such as melittin, succinylated melittin and anionic peptides derived from the N-terminus of HA2 subunit of the influenza virus hemagglutinin, was measured. Melittin (GIGAVLKVLTTGLPALISWIKRKRQQ-NH2) caused a rapid (

Monsigny, M  (1995)

Glycotechnologies- Soft biotechnologies

Biofutur (142) 27-32
Oligosaccharides act as recognition signals as well as effectors both in plants (Oligosaccharins) and in animals. They specifically bind lectins, glycotransferases, glycosidases, adhesion molecules, etc... Membrane lectins are involved in cell-cell recognition phenomena both in normal and in pathological situations. For instance, in case of inflammation the interaction of neutrophils with the endothelial cells is mediated by membrane lectins expressed on the surface of the later cells lining the vessels ; the addition of specific oligosides prevents this interaction and as a consequence the tissue degradation occuring during a prolonged inflammation. Analogous situations are also found in the case of autoimmune diseases and atherosclerosis. Lectins are also involved in recognition between microorganisms (parasite, bacteria, virus) and host cells. Specific oligosaccharides are able to impair this early step of the deleterious processes mediated by microorganisms.

Oligosaccharides act as recognition signals as well as effectors both in plants (Oligosaccharins) and in animals. They specifically bind lectins, glycotransferases, glycosidases, adhesion molecules, etc... Membrane lectins are involved in cell-cell recognition phenomena both in normal and in pathological situations. For instance, in case of inflammation the interaction of neutrophils with the endothelial cells is mediated by membrane lectins expressed on the surface of the later cells lining the vessels ; the addition of specific oligosides prevents this interaction and as a consequence the tissue degradation occuring during a prolonged inflammation. Analogous situations are also found in the case of autoimmune diseases and atherosclerosis. Lectins are also involved in recognition between microorganisms (parasite, bacteria, virus) and host cells. Specific oligosaccharides are able to impair this early step of the deleterious processes mediated by microorganisms.

Gaudin, JC ; Monsigny, M ; Legrand, A  (1995)

Cloning of the CDNA-encoding rabbit galectin-3

Gene 163 (2) 249-252
The complete coding sequence of the rabbit galactin-3-encoding cDNA (LGALS3) has been cloned in a single step by using RT-PCR and specific human LGALS3 cDNA primers. The putative protein contains three domains with different degrees of homology to other known LGALS3. The homology is high in the C-terminal moiety corresponding to the carbohydrate-binding domain and is relatively low in the N-terminal moiety.

The complete coding sequence of the rabbit galactin-3-encoding cDNA (LGALS3) has been cloned in a single step by using RT-PCR and specific human LGALS3 cDNA primers. The putative protein contains three domains with different degrees of homology to other known LGALS3. The homology is high in the C-terminal moiety corresponding to the carbohydrate-binding domain and is relatively low in the N-terminal moiety.


1994   Références trouvées : 14

Sdiqui, N ; Santambien, P ; Roche, AC ; Hebert, E ; Girot, P ; Cochet, S ; Boschetti, E ; Monsigny, M ; Bertrand, O  (1994)

Toxicity studies on native procion-red he-3b and released dye from affinity material exposed to degradative chemical conditions

Journal of Biochemical and Biophysical Methods 29 (3-4) 269-282
Leached ligands from chromatographic packing material submitted to drastic regeneration conditions can contaminate pure biological preparations. These contaminants could have adverse effects from a toxicology point of view that are very poorly documented in liquid chromatography for protein separation. Investigations on toxicity level have been made on released material from immobilized Procion Red HE-3B, after formal identification of the nature of the leached chemical material. Toxicity investigations in vitro involved a number of tests on living cells (eucaryotic and procaryotic) covering different aspects. Behaviour of cells in regular cultures, polyploidia induction, genotoxicity as well as mechanisms of endocytosis have been studied. Results showed no toxic effects within the range of concentration of dye and dye derivatives studied. Genotoxicity studies in particular did not show any toxic effect over a range of concentration much higher than the regular level of dye leakage from the sorbent.

Leached ligands from chromatographic packing material submitted to drastic regeneration conditions can contaminate pure biological preparations. These contaminants could have adverse effects from a toxicology point of view that are very poorly documented in liquid chromatography for protein separation. Investigations on toxicity level have been made on released material from immobilized Procion Red HE-3B, after formal identification of the nature of the leached chemical material. Toxicity investigations in vitro involved a number of tests on living cells (eucaryotic and procaryotic) covering different aspects. Behaviour of cells in regular cultures, polyploidia induction, genotoxicity as well as mechanisms of endocytosis have been studied. Results showed no toxic effects within the range of concentration of dye and dye derivatives studied. Genotoxicity studies in particular did not show any toxic effect over a range of concentration much higher than the regular level of dye leakage from the sorbent.

Kuchler, S ; Graff, MN ; Gobaille, S ; Vincendon, G ; Roche, AC ; Delaunoy, JP ; Monsigny, M ; Zanetta, JP  (1994)

Mannose dependent tightening of the rat ependymal cell barrier - in vivo and in vitro study using neoglycoproteins

Neurochemistry International 24 (1) 43-55
The possible role of carbohydrate binding proteins (lectins) and glycoconjugates in the formation of junctions ensuring tightening between ependymal cells was studied using synthetic glycoconjugates, the neoglycoproteins. These compounds are prepared by substituting bovine serum albumin with sugar residues and additional labelling (or not) with fluorescein or biotin. Injections of these components into the cerebral ventricles of adult rats resulted in a binding pattern which could be related to their carbohydrate composition. Mannose-containing neoglycoproteins were bound to ependymal cell cilia and penetrated rapidly the brain tissue. Such phenomenon was not seen with glucose- or galactose-containing neoglycoprotein molecules. In contrast, mannose-, galactose- and glucose- containing neoglycoproteins bound strongly to some endothelial cells around blood vessels. Fluorescent unglycosylated serum albumin did not bind to any brain structures.

The possible role of carbohydrate binding proteins (lectins) and glycoconjugates in the formation of junctions ensuring tightening between ependymal cells was studied using synthetic glycoconjugates, the neoglycoproteins. These compounds are prepared by substituting bovine serum albumin with sugar residues and additional labelling (or not) with fluorescein or biotin. Injections of these components into the cerebral ventricles of adult rats resulted in a binding pattern which could be related to their carbohydrate composition. Mannose-containing neoglycoproteins were bound to ependymal cell cilia and penetrated rapidly the brain tissue. Such phenomenon was not seen with glucose- or galactose-containing neoglycoprotein molecules. In contrast, mannose-, galactose- and glucose- containing neoglycoproteins bound strongly to some endothelial cells around blood vessels. Fluorescent unglycosylated serum albumin did not bind to any brain structures.

Monsigny, M ; Roche, AC ; Midoux, P ; Mayer, R  (1994)

Glycoconjugates as carriers for specific delivery of therapeutic drugs and genes

Advanced Drug Delivery Reviews 14 (1) 1-24
Cell surface receptors are good candidates to selectively target drugs, oligonucleotides or even genes by making use of their specific ligands. A large number of mammalian cells express cell surface sugar-binding proteins, also called ''membrane lectins''. Therefore, sugars may be used as specific recognition signals to specifically deliver biological active components. Tens of membrane lectins with different sugar specificities have been characterized ; some of them actively carry their ligands to intracellular compartments, including endosomes, lysosomes and, in some cases, Golgi apparatus.

Cell surface receptors are good candidates to selectively target drugs, oligonucleotides or even genes by making use of their specific ligands. A large number of mammalian cells express cell surface sugar-binding proteins, also called ’’membrane lectins’’. Therefore, sugars may be used as specific recognition signals to specifically deliver biological active components. Tens of membrane lectins with different sugar specificities have been characterized ; some of them actively carry their ligands to intracellular compartments, including endosomes, lysosomes and, in some cases, Golgi apparatus.

Carpentier, V ; Vassard, C ; plessis, C ; Motta, G ; Monsigny, M ; Roche, AC  (1994)

Characterization and cellular-localization by monoclonal-antibodies of the 60-kda mannose-specific lectin of human promyelocytic cells, hl-60

Glycoconjugate Journal 11 (4) 333-338
Myelomonocytic lineage cells express an M(r) 60000 mannose specific lectin, MR60 (Pimpaneau et al. (1991), Carbohydr Res 213 : 95-108). Under non-reducing conditions, this protein migrates as a 120000 protein. MR60 does not contain any N-glycan moiety cleavable by the action of N-glycanase. MR60 induces a sugar selective aggregation of beads coated with glycosylated albumin : beads bearing a-D-mannosyl residues are aggregated while beads bearing a-D-glucosyl residues are not. A monoclonal antibody Lec101B, specific for MR60, recognizes a single M(r) 60000 protein by Western blotting. This monoclonal antibody does not label the cell surface of cells expressing MR60, but decorates intracellular vesicles upon permeabilization of these cells.

Myelomonocytic lineage cells express an M(r) 60000 mannose specific lectin, MR60 (Pimpaneau et al. (1991), Carbohydr Res 213 : 95-108). Under non-reducing conditions, this protein migrates as a 120000 protein. MR60 does not contain any N-glycan moiety cleavable by the action of N-glycanase. MR60 induces a sugar selective aggregation of beads coated with glycosylated albumin : beads bearing a-D-mannosyl residues are aggregated while beads bearing a-D-glucosyl residues are not. A monoclonal antibody Lec101B, specific for MR60, recognizes a single M(r) 60000 protein by Western blotting. This monoclonal antibody does not label the cell surface of cells expressing MR60, but decorates intracellular vesicles upon permeabilization of these cells.

Bertrand, O. ; Boschetti, E. ; Cochet, S. ; Girot, P. ; Hebert, E. ; Monsigny, M. ; Roche, A.-C. ; Santambien, P. ; Sdiqui, N.  (1994)

Toxicity studies on Reactive Blue-2 leached from affinity material exposed to extreme chemical conditions

Bioseparation 4 (5) 299-309
Toxicity effects related to leached ligands from affinity sorbents that can contaminate biological preparations were investigated in the particular case of immobilized Reactive Blue-2. Initially, identification of the real chemical structure of leached dye has been done by HPLC after incubation in extreme conditions. Toxicity investigations in vitro involving several well known tests showed no toxic effects within the studied range of dye concentration. Cell cultures behaved normally when the adhesion phase was successful ; polyploidy induction in human cells by the native dye and its derivatives identified as possible leached material was very similar to standard cultures. Genotoxicity studies did not evidence any toxic effect in E. coli cultures of dyes themselves or of the same dyes after metabolic activation.

Toxicity effects related to leached ligands from affinity sorbents that can contaminate biological preparations were investigated in the particular case of immobilized Reactive Blue-2. Initially, identification of the real chemical structure of leached dye has been done by HPLC after incubation in extreme conditions. Toxicity investigations in vitro involving several well known tests showed no toxic effects within the studied range of dye concentration. Cell cultures behaved normally when the adhesion phase was successful ; polyploidy induction in human cells by the native dye and its derivatives identified as possible leached material was very similar to standard cultures. Genotoxicity studies did not evidence any toxic effect in E. coli cultures of dyes themselves or of the same dyes after metabolic activation.

Wisniewski, JP ; Monsigny, M ; Delmotte, FM  (1994)

Purification of an a-L-fucoside-binding protein from rhizobium-lupini

Biochimie 76 (2) 121-128
Lectins associated with the bacterial cell surface of Rhizobium lupini strain LL13 were evidenced by erythrocyte agglutination, by aggregation of neoglycoprotein coated beads and by spectrofluorimetry using fluoresceinylated neoglycoproteins. At pH 5.0, a specific binding of the fluorescein-labelled neoglycoprotein bearing a-L-fucose was observed. The binding of this labelled neoglycoprotein is a saturable phenomenon and is inhibited by the same unlabelled neoglycoprotein. Extracts of R lupini obtained by disrupting a bacterial pellet through a French press were stabilized at pH 5.6 by gel filtration and purified to homogeneity by affinity chromatography on Agarose A4 substituted with a-L-fucose. A protein with a M(r) approximate to 19 000 was specifically eluted from this affinity column with L-fucose. Isoelectric focusing of this sample yielded a single band with pi near 6.7. This protein specifically aggregated L-Fuc-BSA-coated microspheres. The results obtained in the present study indicate that we have purified from Rhizobium lupini strain LL13, a L-fucose binding protein as a lectin.

Lectins associated with the bacterial cell surface of Rhizobium lupini strain LL13 were evidenced by erythrocyte agglutination, by aggregation of neoglycoprotein coated beads and by spectrofluorimetry using fluoresceinylated neoglycoproteins. At pH 5.0, a specific binding of the fluorescein-labelled neoglycoprotein bearing a-L-fucose was observed. The binding of this labelled neoglycoprotein is a saturable phenomenon and is inhibited by the same unlabelled neoglycoprotein. Extracts of R lupini obtained by disrupting a bacterial pellet through a French press were stabilized at pH 5.6 by gel filtration and purified to homogeneity by affinity chromatography on Agarose A4 substituted with a-L-fucose. A protein with a M(r) approximate to 19 000 was specifically eluted from this affinity column with L-fucose. Isoelectric focusing of this sample yielded a single band with pi near 6.7. This protein specifically aggregated L-Fuc-BSA-coated microspheres. The results obtained in the present study indicate that we have purified from Rhizobium lupini strain LL13, a L-fucose binding protein as a lectin.

Florent, I ; Lebonniec, S ; Carcy, B ; Grellier, P ; Mercerau-Puijalon, O ; Bonnefoy, S ; Dhermy, D ; Monsigny, M ; Mayer, R ; Schrevel, J  (1994)

Plasmodium-falciparum proteinases - cloning of the putative gene coding for the merozoite proteinase for erythrocyte invasion (MPEI) and determination of hydrolysis sites of spectrin by PF37 proteinase

Memorias Do Instituto Oswaldo Cruz 8947-49 Suppl. 2
Numerous proteinase activities have been shown to be essential for the survival of Plasmodium falciparum. One approach to antimalarial chemotherapy, would be to block specially one or several of these activities, by using compounds structurally analogous to the substrates of these proteinases. Such a strategy requires a detailed knowledge of the active site of the proteinase, in order to identify the best substrate for the proteinase. Aiming at developing such a strategy, two proteinases previously identified in our laboratory were chosen for further characterization of their molecular structure and properties : the merozoite proteinase for erythrocytic invasion (MPEI), involved in the erythrocyte invasion by the merozoites, and the Pf37 proteinase, which hydrolyses human spectrin in vitro.

Numerous proteinase activities have been shown to be essential for the survival of Plasmodium falciparum. One approach to antimalarial chemotherapy, would be to block specially one or several of these activities, by using compounds structurally analogous to the substrates of these proteinases. Such a strategy requires a detailed knowledge of the active site of the proteinase, in order to identify the best substrate for the proteinase. Aiming at developing such a strategy, two proteinases previously identified in our laboratory were chosen for further characterization of their molecular structure and properties : the merozoite proteinase for erythrocytic invasion (MPEI), involved in the erythrocyte invasion by the merozoites, and the Pf37 proteinase, which hydrolyses human spectrin in vitro.

Santambien, P. ; Sdiqui, N. ; Hebert, E. ; Girot, P. ; Hulak, I. ; Roche, A.-C. ; Monsigny, M. ; Boschetti, E.  (1994)

In vitro and in vivo toxicity assays for dye ligands used in affinity chromatography

Annales Pharmaceutiques Francaises 52 (3) 137-152
Some reactive textile dyes are used for years as biomimetic ligands in protein purification. The reluctance to use these performant systems in large scale for therapeutically applicable proteins is related with the possible dye leakage and consequently with problems of contamination. Therefore, toxicology data are necessary to qualify the level of danger in association with sensitive assays. This study deals with a series of in vitro toxicity studies with eucaryotic cells (growth, polyploidia, ...) as well as with procaryotic cells (E. coli) for genotoxic studies. Both approaches demonstrated a total absence of toxicity for all ranges of concentrations investigated for Reactive Blue 2, Reactive Red 120 and their derivatives. Additional experiments done in vivo by the administration of dye solutions to a series of mice confirmed the non toxic character of these dyes in vitro.

Some reactive textile dyes are used for years as biomimetic ligands in protein purification. The reluctance to use these performant systems in large scale for therapeutically applicable proteins is related with the possible dye leakage and consequently with problems of contamination. Therefore, toxicology data are necessary to qualify the level of danger in association with sensitive assays. This study deals with a series of in vitro toxicity studies with eucaryotic cells (growth, polyploidia, ...) as well as with procaryotic cells (E. coli) for genotoxic studies. Both approaches demonstrated a total absence of toxicity for all ranges of concentrations investigated for Reactive Blue 2, Reactive Red 120 and their derivatives. Additional experiments done in vivo by the administration of dye solutions to a series of mice confirmed the non toxic character of these dyes in vitro.

Barondes, SH ; Castronovo, V ; Cooper, DNW ; Cummings, RD ; Drickamer, K ; Feizi, T ; Gitt, MA ; Hirabayashi, J ; Hughes, C ; Kasai, K ; Leffler, H ; Liu, FT ; Lotan, R ; Mercurio, AM ; Monsigny, M ; Pillai, S ; Poirer, F ; Raz, A ; Rigby, PWJ ; Rini, JM ; Wang, JL  (1994)

Galectins - a family of animal ß-galactoside-binding lectins

Cell 76 (4) 597-598

Hebert, E ; Monsigny, M  (1994)

Galectin-3 messenger-RNA level depends on transformation phenotype in RAS-transformed NIH 3T3 cells

Biology of The Cell 81 (1) 73-76
The increase in galectin-3 lectin content observed in tumours or in in vitro transformed cells suggests that this lectin is important in the transformation process. In the present study, we investigated the mRNA expression level of the galectin-3, galectin-1 and macrophage mannose receptor in normal and ras-transformed NIH 3T3 cells in relation to their transformation state. The galectin-3 mRNA content in ras-transformed cells is increased in fully transformed cells, with a maximum in ras-transformed cells that have lost their growth anchorage-dependence. Under the same conditions, the galectin-1 mRNA level which was high in normal cells, increased slightly in transformed cells. The mRNA for the macrophage mannose receptor was not detected in 3T3 cells or in their ras-transformed counterparts.

The increase in galectin-3 lectin content observed in tumours or in in vitro transformed cells suggests that this lectin is important in the transformation process. In the present study, we investigated the mRNA expression level of the galectin-3, galectin-1 and macrophage mannose receptor in normal and ras-transformed NIH 3T3 cells in relation to their transformation state. The galectin-3 mRNA content in ras-transformed cells is increased in fully transformed cells, with a maximum in ras-transformed cells that have lost their growth anchorage-dependence. Under the same conditions, the galectin-1 mRNA level which was high in normal cells, increased slightly in transformed cells. The mRNA for the macrophage mannose receptor was not detected in 3T3 cells or in their ras-transformed counterparts.

Monsigny, M  (1994)

Endogenous glycoproteins and lectins, a sweet interplay

M S-Medecine Sciences 10 (1) 9-11

Chevrie, K ; Motta, G ; Mayer, R ; Monsigny, M  (1994)

Dye-hydrophobic hapten conjugate anti-dye antibody complex as immunogen - preparation of hydrophobic hapten-specific monoclonal-antibodies

Biochimie 76 (2) 171-179
In order to induce the production of antibodies specific for small molecules, it is common to link them to a protein. However, when the small molecule is very hydrophobic it is extremely difficult to prepare such a conjugate. Here, we describe a simple way to obtain an antigenic conjugate under controlled conditions : in a first step a very hydrophobic hapten, cholanic acid, is linked to a dye, basilen blue, in organic solvent ; in a second step the cholanic acid-basilen blue conjugate is dissolved in phosphate buffered saline and mixed with rabbit polyclonal anti-basilen blue antibodies previously raised in rabbits against basilen blue-keyhole limpet hemocyanin conjugate. Such a complex, which dissociates very slowly, appears to be a good immunogen in mice. Anti-cholanyl residue monoclonal antibodies were produced and characterized.

In order to induce the production of antibodies specific for small molecules, it is common to link them to a protein. However, when the small molecule is very hydrophobic it is extremely difficult to prepare such a conjugate. Here, we describe a simple way to obtain an antigenic conjugate under controlled conditions : in a first step a very hydrophobic hapten, cholanic acid, is linked to a dye, basilen blue, in organic solvent ; in a second step the cholanic acid-basilen blue conjugate is dissolved in phosphate buffered saline and mixed with rabbit polyclonal anti-basilen blue antibodies previously raised in rabbits against basilen blue-keyhole limpet hemocyanin conjugate. Such a complex, which dissociates very slowly, appears to be a good immunogen in mice. Anti-cholanyl residue monoclonal antibodies were produced and characterized.

Rouleux, F ; Monsigny, M ; Legrand, A  (1994)

A negative regulatory element of the macrophage-specific human mannose receptor gene represses its expression in nonmyeloid cells

Experimental Cell Research 214 (1) 113-119
We have cloned the putative promoter of the human mannose receptor gene using the ligation-mediated polymerase chain reaction. This modified polymerase chain reaction method depends upon the ligation of restricted genomic DNA fragments to a sequence of DNA containing a generic primer site. Approximately 400 bp of genomic DNA sequence immediately upstream from the 5' end of the lectin gene was amplified with this strategy. Primer-extended reverse transcription identified several 5' ends of the mannose receptor mRNA corresponding to differential use of initiation transcription sites.

We have cloned the putative promoter of the human mannose receptor gene using the ligation-mediated polymerase chain reaction. This modified polymerase chain reaction method depends upon the ligation of restricted genomic DNA fragments to a sequence of DNA containing a generic primer site. Approximately 400 bp of genomic DNA sequence immediately upstream from the 5’ end of the lectin gene was amplified with this strategy. Primer-extended reverse transcription identified several 5’ ends of the mannose receptor mRNA corresponding to differential use of initiation transcription sites.

Bizouarne, N ; Denis, V ; Legrand, A ; Monsigny, M ; Kieda, C  (1994)

A SV40 immortalized murine endothelial-cell line from peripheral lymph-node high endothelium expresses a new a-L-fucose binding-protein

Biology of The Cell 79 (3) 209-218
Endothelial cells from mouse peripheral lymph nodes were immortalized by cationic liposome-mediated transfection using a plasmid construct containing both the gene coding for the large T antigen of simian virus 40 and a geneticin resistance gene suitable for selection. A cell line (HECa10) was isolated on the basis of its capacity to specifically bind fucoside carrying glycoconjugates ; these cells present the main characteristics of endothelial cells : production of angiotensin converting enzyme and of factor VIII-related antigen.

Endothelial cells from mouse peripheral lymph nodes were immortalized by cationic liposome-mediated transfection using a plasmid construct containing both the gene coding for the large T antigen of simian virus 40 and a geneticin resistance gene suitable for selection. A cell line (HECa10) was isolated on the basis of its capacity to specifically bind fucoside carrying glycoconjugates ; these cells present the main characteristics of endothelial cells : production of angiotensin converting enzyme and of factor VIII-related antigen.


1993   Références trouvées : 16

Duverger, E ; Carpentier, V ; Roche, AC ; Monsigny, M  (1993)

Sugar-dependent nuclear import of glycoconjugates from the cytosol

Experimental Cell Research 207 (1) 197-201

Monsigny, M  (1993)

Transgenesis in glycobiology - role of the lectin - P-selectin

M S-Medecine Sciences 9 (11) 1270-1271

Monsigny, M ; Midoux, P ; Roche, AC  (1993)

Targeted plasmid-polylysine complex as putative tools to selectively transfer genes ex vivo and in vivo

M S-Medecine Sciences 9 (4) 441-449
Gene therapy will be an ideal method when it is possible to transfer DNA in cells with a great efficacy and an absolute safety in vivo. Along with viral carriers which are efficient but may not be strictly safe, non viral carriers could be advantageously used. Polylysine, a polycation giving stable DNA complexes, is proposed as the basis of such a carrier system. Indeed, polylysine substituted with either a protein easily taken up by cells due to specific cell surface receptors, or carbohydrate moieties which selectively interact with lectins expressed at the surface of specific cells, allows a cell specific delivery of genes. In addition, when such a targeted carrier is used together with components that protect the plasmid-targeted polylysine complex from the hydrolytic activities of lysosomes and/or help the complex to cross the cell membranes to reach the cytosol, the specificity and the efficacy of the transfection are conspicuously higher.

Gene therapy will be an ideal method when it is possible to transfer DNA in cells with a great efficacy and an absolute safety in vivo. Along with viral carriers which are efficient but may not be strictly safe, non viral carriers could be advantageously used. Polylysine, a polycation giving stable DNA complexes, is proposed as the basis of such a carrier system. Indeed, polylysine substituted with either a protein easily taken up by cells due to specific cell surface receptors, or carbohydrate moieties which selectively interact with lectins expressed at the surface of specific cells, allows a cell specific delivery of genes. In addition, when such a targeted carrier is used together with components that protect the plasmid-targeted polylysine complex from the hydrolytic activities of lysosomes and/or help the complex to cross the cell membranes to reach the cytosol, the specificity and the efficacy of the transfection are conspicuously higher.

Arar, K ; Monsigny, M ; Mayer, R  (1993)

Synthesis of oligonucleotide-peptide conjugates containing a kdel signal sequence

Tetrahedron Letters 34 (50) 8087-8090
An improved method of preparation of oligonucleotide-peptide conjugates is described. An oligopeptide containing a and epsilon-aminogroups is mainly substituted at its a-NH2 end by epsilon-maleimidocaproic acid-N-hydroxysuccinimide ester at pH 6.5 for 1 h. The N-a-maleimidocaproyl-peptide derivative, purified by HPLC, reacts with the thiol group of an oligonucleotide in a 82% yield at pH 7.2. The thiol group is generated in situ by the action of tris(carboxyethyl)phosphine on an oligonucleotide bearing a disulfide bridge.

An improved method of preparation of oligonucleotide-peptide conjugates is described. An oligopeptide containing a and epsilon-aminogroups is mainly substituted at its a-NH2 end by epsilon-maleimidocaproic acid-N-hydroxysuccinimide ester at pH 6.5 for 1 h. The N-a-maleimidocaproyl-peptide derivative, purified by HPLC, reacts with the thiol group of an oligonucleotide in a 82% yield at pH 7.2. The thiol group is generated in situ by the action of tris(carboxyethyl)phosphine on an oligonucleotide bearing a disulfide bridge.

Negre, E ; Monsigny, M ; Mayer, R  (1993)

Synthesis of an allopurinol riboside-mannosylated poly-L-lysine conjugate

Tetrahedron 49 (32) 6991-7000
The synthesis of a mannosylated carrier used as a drug delivery system to target specifically an antileishmanial drug allopurinol riboside [4-hydroxy-1-ß-D-ribofuranosyl-1H-pyrazolo(3,4-d) pyrimidine] into Leishmania donovani infected macrophages via their membrane lectin is described. The synthetic construct is made of a poly-L-lysine backbone, partially acylated with d-gluconolactone to enhance its water-solubility and substituted with glycyl-glycine asa spacer arm. O-p-phenylisothiocyanate-5'-phosphodiester derivatives of allopurinol riboside and of inosine were synthesized and attached to the spacer arm, then the remaining spacer arms were substituted by reaction with phenyl acetate mannosyl residues.

The synthesis of a mannosylated carrier used as a drug delivery system to target specifically an antileishmanial drug allopurinol riboside [4-hydroxy-1-ß-D-ribofuranosyl-1H-pyrazolo(3,4-d) pyrimidine] into Leishmania donovani infected macrophages via their membrane lectin is described. The synthetic construct is made of a poly-L-lysine backbone, partially acylated with d-gluconolactone to enhance its water-solubility and substituted with glycyl-glycine asa spacer arm. O-p-phenylisothiocyanate-5’-phosphodiester derivatives of allopurinol riboside and of inosine were synthesized and attached to the spacer arm, then the remaining spacer arms were substituted by reaction with phenyl acetate mannosyl residues.

Midoux, P ; Mendes, C ; Legrand, A ; Raimond, J ; Mayer, R ; Monsigny, M ; Roche, AC  (1993)

Specific gene-transfer mediated by lactosylated poly-Ll-lysine into hepatoma-cells

Nucleic Acids Research 21 (4) 871-878
Plasmid DNA/glycosylated polylysine complexes were used to transfer in vitro a luciferase reporter gene into human hepatoma cells by a receptor-mediated endocytosis process. HepG2 cells which express a galactose specific membrane lectin were efficiently and selectively transfected with pSV2Luc/lactosylated polylysine complexes in a sugar dependent manner :

Plasmid DNA/glycosylated polylysine complexes were used to transfer in vitro a luciferase reporter gene into human hepatoma cells by a receptor-mediated endocytosis process. HepG2 cells which express a galactose specific membrane lectin were efficiently and selectively transfected with pSV2Luc/lactosylated polylysine complexes in a sugar dependent manner :

Anjuere, F ; Monsigny, M ; Lelievre, Y ; Mayer, R  (1993)

Sensitive, hydrosoluble, macromolecular fluorogenic substrates for human immunodeficiency virus-1 proteinase

Biochemical Journal 291 869-873 Part 3
Hydrosoluble macromolecular fluorogenic substrates specific for the human immunodeficiency virus 1 (HIV-1) proteinase have been prepared. The fluoresceinyl peptide Ftc-epsilon-Ahx-Ser-Phe-Asn-Phe-Pro-Gln-Ile-Thr-(Gly)n, corresponding to the first cleavage site of HIV-1 gag-pol native precursor was linked to a water-soluble neutral (Lys)n derivative. The epsilon-aminohexanoyl residue (epsilon-Ahx) and the glycyl sequence were added in order to improve the stability of the substrate and the accessibility of the cleavage site to the HIV-1 proteinase respectively. This macromolecular peptidic-substrate conjugate is significantly more water-soluble than the free peptide itself on a substrate molar concentration basis.

Hydrosoluble macromolecular fluorogenic substrates specific for the human immunodeficiency virus 1 (HIV-1) proteinase have been prepared. The fluoresceinyl peptide Ftc-epsilon-Ahx-Ser-Phe-Asn-Phe-Pro-Gln-Ile-Thr-(Gly)n, corresponding to the first cleavage site of HIV-1 gag-pol native precursor was linked to a water-soluble neutral (Lys)n derivative. The epsilon-aminohexanoyl residue (epsilon-Ahx) and the glycyl sequence were added in order to improve the stability of the substrate and the accessibility of the cleavage site to the HIV-1 proteinase respectively. This macromolecular peptidic-substrate conjugate is significantly more water-soluble than the free peptide itself on a substrate molar concentration basis.

Schrevel, J. ; Barrault, C. ; Deguercy, A. ; Grellier, P. ; Lawton, P. ; Heidrich, H.G. ; Caballero, M. ; Monsigny, M. ; Mayer, R.  (1993)

Plasmodium falciparum proteinases and red blood cell invasion.

Parassitologia (Rome) 35 103-105

Hebert, E ; Monsigny, M  (1993)

Oncogenes and expression of endogenous lectins and glycoconjugates

Biology of The Cell 79 (2) 97-109
It is now well established that malignant transformation of eucaryotic cells is concomitant with typical alterations of glycosylation and the expression pattern of endogenous lectins. In parallel, oncogene transfection studies revealed a correlation between the expression of some of these genes, the transformed state and perhaps metastasis. These observations lead to the idea that oncogenes may control the expression of enzymes involved in the biosynthetic pathway of cell membrane glycoconjugates and the expression of endogenous lectins. Indeed, several contributions have shown that cells upon transfection with activated oncogenes of the ras family become invasive and/or metastatic and have their membrane glycoproteins modified. Information on the molecular mechanism of this postulated oncogene regulation is still lacking. Because of the diversity of the functions of oncogene-encoded proteins, further experiments dealing with other activated oncogenes may help in deciphering the regulation of expression of glycoconjugates and endogenous lectins together with their functions.

It is now well established that malignant transformation of eucaryotic cells is concomitant with typical alterations of glycosylation and the expression pattern of endogenous lectins. In parallel, oncogene transfection studies revealed a correlation between the expression of some of these genes, the transformed state and perhaps metastasis. These observations lead to the idea that oncogenes may control the expression of enzymes involved in the biosynthetic pathway of cell membrane glycoconjugates and the expression of endogenous lectins. Indeed, several contributions have shown that cells upon transfection with activated oncogenes of the ras family become invasive and/or metastatic and have their membrane glycoproteins modified. Information on the molecular mechanism of this postulated oncogene regulation is still lacking. Because of the diversity of the functions of oncogene-encoded proteins, further experiments dealing with other activated oncogenes may help in deciphering the regulation of expression of glycoconjugates and endogenous lectins together with their functions.

Elmoudni, B ; Philippe, M ; Monsigny, M ; Schrevel, J  (1993)

N-acetylglucosamine-binding proteins on plasmodium-falciparum merozoite surface

Glycobiology 3 (4) 305-312
Plasmodium falciparum merozoite surface is specifically labelled with a neoglycoprotein bearing N-acetylglucosamine (GlcNAc) residues in a sugar-dependent manner, as shown by affinity cytochemistry in fluorescence and electron microscopy. To ascertain the nature of the sugar receptor, merozoite proteins were blotted and tested by a two-step method using biotinylated GlcNAc-bovine serum albumin (BSA) and streptavidin-peroxidase conjugate. Three parasite proteins were specifically revealed and designated as Pf 120, Pf 83 and Pf 45 GlcNAc-binding proteins. These proteins bind to a gel substituted with GlcNAc and are specifically eluted with 300 mM GlcNAc.

Plasmodium falciparum merozoite surface is specifically labelled with a neoglycoprotein bearing N-acetylglucosamine (GlcNAc) residues in a sugar-dependent manner, as shown by affinity cytochemistry in fluorescence and electron microscopy. To ascertain the nature of the sugar receptor, merozoite proteins were blotted and tested by a two-step method using biotinylated GlcNAc-bovine serum albumin (BSA) and streptavidin-peroxidase conjugate. Three parasite proteins were specifically revealed and designated as Pf 120, Pf 83 and Pf 45 GlcNAc-binding proteins. These proteins bind to a gel substituted with GlcNAc and are specifically eluted with 300 mM GlcNAc.

Kieda, C ; Bizouarne, N ; Denis, V ; Monsigny, M  (1993)

Lymphocyte/endothelial cell recognitions - involvement of a new fucose-binding protein

Journal of Cellular Biochemistry 356-356 Suppl. 17a

Kieda, C ; Denis, V ; Monsigny, M  (1993)

Lectin-glycoconjugate recognition in lymphocyte-endothelial cell-interactions

Glycoconjugate Journal 10 (4) 268-268

Monsigny, M  (1993)

Glycoconjugates - recognition process in animals

Biofutur 125 20-24

Carpentier, V ; Midoux, P ; Monsigny, M ; Roche, AC  (1993)

Endocytosis of a(1)-acid glycoprotein variants by human monocytic lineage cells

Biology of The Cell 77 (2) 187-193
Human a1-acid glycoprotein (AGP or orosomucoid) is a major glycoprotein of plasma. AGP can be separated on immobilized concanavalin A into three variants bearing none (AGP A), one (AGP B) or two (AGP C) biantennary glycans. In this paper, we show, using flow cytometry and confocal microscopy, that AGP C which is eluted from concanavalin A with mannose, binds to human monocytes, monocyte-derived macrophages as well as human promonocytic cell lines such as THP1 or U937. Conversely HL60, a promyelocytic cell line, does not express the surface AGP C binding protein. AGP C is internalized and degraded with an efficiency depending on the state of differentiation of these cells. In contrast, AGP A which is not recognized by concanavalin A, does not bind to any of these cells.

Human a1-acid glycoprotein (AGP or orosomucoid) is a major glycoprotein of plasma. AGP can be separated on immobilized concanavalin A into three variants bearing none (AGP A), one (AGP B) or two (AGP C) biantennary glycans. In this paper, we show, using flow cytometry and confocal microscopy, that AGP C which is eluted from concanavalin A with mannose, binds to human monocytes, monocyte-derived macrophages as well as human promonocytic cell lines such as THP1 or U937. Conversely HL60, a promyelocytic cell line, does not express the surface AGP C binding protein. AGP C is internalized and degraded with an efficiency depending on the state of differentiation of these cells. In contrast, AGP A which is not recognized by concanavalin A, does not bind to any of these cells.

Bizouarne, N ; Mitterrand, M ; Monsigny, M ; Kieda, C  (1993)

Characterization of membrane sugar-specific receptors in cultured high endothelial-cells from mouse peripheral lymph-nodes

Biology of The Cell 79 (1) 27-35
The culture of specialized high endothelial cells (HEC) from lymphoid organs (peripheral lymph nodes (PLN) and Peyer's patches (PP)) was undertaken in order to study and characterize the cell surface molecules which are involved in lymphocyte recognition and allow homing. Cells were stimulated in vivo by a graft versus host (GVH) type of reaction before isolation and culture. The resulting adherent and growing cells were characterized as endothelial cells because of their typical aspect and their ability to produce angiotensin-converting enzyme and factor VIII-related antigen. They possess tissue-specific endothelial addressins. MECA 79 antigen is present on cells isolated from PLN while MECA 367 antigen is detected on cells from PP.

The culture of specialized high endothelial cells (HEC) from lymphoid organs (peripheral lymph nodes (PLN) and Peyer’s patches (PP)) was undertaken in order to study and characterize the cell surface molecules which are involved in lymphocyte recognition and allow homing. Cells were stimulated in vivo by a graft versus host (GVH) type of reaction before isolation and culture. The resulting adherent and growing cells were characterized as endothelial cells because of their typical aspect and their ability to produce angiotensin-converting enzyme and factor VIII-related antigen. They possess tissue-specific endothelial addressins. MECA 79 antigen is present on cells isolated from PLN while MECA 367 antigen is detected on cells from PP.

Monsigny, M  (1993)

An efficient drug against influenza - is it for tomorrow ?

M S-Medecine Sciences 9 (11) 1274-1274


1992   Références trouvées : 9

Bonfils, E ; Mendes, C ; Roche, AC ; Monsigny, M ; Midoux, P  (1992)

Uptake by macrophages of a biotinylated oligo- ?-deoxythymidylate by using mannosylated streptavidin

Bioconjugate Chemistry 3 (4) 277-284
Streptavidin substituted with mannose residues increased by 20-fold the intracellular concentration of a biotinylated dodecakis(a-deoxythymidylate) in macrophages by comparison with the uptake of free oligodeoxynucleotide. Streptavidin, the bacterial homologue of the very basic avidin, which does not contain any carbohydrate moieties and is a neutral protein, was substituted with 12 mannose residues in order to be recognized and internalized by mannose-specific lectins on the surface of macrophages. A 3'-biotinylated and 5'-fluoresceinylated dodecakis(a-deoxythymidylate) was synthesized and bound onto mannosylated streptavidin. The conjugate was isolated, and by using flow cytometry, it was shown that the uptake of fluoresceinylated oligodeoxynucleotides bound to mannosylated streptavidin by macrophages is 20-fold higher than that of free oligodeoxynucleotides and that the uptake was competively inhibited by mannosylated serum albumin. Glycosylated streptavidin conjugates recognizing specific membrane lectins on different cells provide the possibility to target biotinylated antisense oligodeoxynucleotides and to increase the biological effect of these chemotherapeutic agents.

Streptavidin substituted with mannose residues increased by 20-fold the intracellular concentration of a biotinylated dodecakis(a-deoxythymidylate) in macrophages by comparison with the uptake of free oligodeoxynucleotide. Streptavidin, the bacterial homologue of the very basic avidin, which does not contain any carbohydrate moieties and is a neutral protein, was substituted with 12 mannose residues in order to be recognized and internalized by mannose-specific lectins on the surface of macrophages. A 3’-biotinylated and 5’-fluoresceinylated dodecakis(a-deoxythymidylate) was synthesized and bound onto mannosylated streptavidin. The conjugate was isolated, and by using flow cytometry, it was shown that the uptake of fluoresceinylated oligodeoxynucleotides bound to mannosylated streptavidin by macrophages is 20-fold higher than that of free oligodeoxynucleotides and that the uptake was competively inhibited by mannosylated serum albumin. Glycosylated streptavidin conjugates recognizing specific membrane lectins on different cells provide the possibility to target biotinylated antisense oligodeoxynucleotides and to increase the biological effect of these chemotherapeutic agents.

Breton, P ; Petit, C ; Bousser, MT ; Monsigny, M ; Mayer, R ; Roche, AC  (1992)

Secretion of macrophage cytotoxic factors induced by new desmuramyl acylpseudopeptide analogs of muramyl dipeptide

Oncology Research 4 (3) 103-107
Sixteen desmuramyl analogs of muramyl dipeptide (MDP) were tested for their abilities to stimulate cytotoxic factor secretion by mouse peritoneal macrophages. Among them, the pseudohexapeptide Boc-Gly psi[CH2O]-D-Ala-Ala-D-Glu[Lys(H-Gly)-NHEt]-NH2-appeared to be four times more effective than MDP. From this study, the D configuration of the pseudo-alanyl (or lactyl) residue appears to be essential for activity.

Sixteen desmuramyl analogs of muramyl dipeptide (MDP) were tested for their abilities to stimulate cytotoxic factor secretion by mouse peritoneal macrophages. Among them, the pseudohexapeptide Boc-Gly psi[CH2O]-D-Ala-Ala-D-Glu[Lys(H-Gly)-NHEt]-NH2-appeared to be four times more effective than MDP. From this study, the D configuration of the pseudo-alanyl (or lactyl) residue appears to be essential for activity.

Bonfils, E ; Depierreux, C ; Midoux, P ; Thuong, NT ; Monsigny, M ; Roche, AC  (1992)

Drug targeting - synthesis and endocytosis of oligonucleotide-neoglycoprotein conjugates

Nucleic Acids Research 20 (17) 4621-4629
Inhibition of gene expression by antisense oligonucleotides is limited by their low ability to enter cells. Knowing that sugar binding receptors, also called membrane lectins, efficiently internalize neoglycoproteins bearing the relevant sugar, 6-phosphomannose, for instance, oligonucleotides-substituted on their 5'-end with either a fluorescent probe or a radioactive label on the one hand, and bearing a thiol function on their 3'-end, on the other hand,-were coupled onto 6-phosphomannosylated proteins via a disulfide bridge. The oligonucleotide bound to 6-phosphomannosylated serum albumin is much more efficiently internalized roughly 20 times than the free oligonucleotide. Although most of the oligonucleotides are associated with vesicular compartments, oligonucleotides after releasing from the carrier by reduction of the disulfide bridge may find their way to reach the cytosol and then lead to an increase in the efficiency of the oligonucleotides.

Inhibition of gene expression by antisense oligonucleotides is limited by their low ability to enter cells. Knowing that sugar binding receptors, also called membrane lectins, efficiently internalize neoglycoproteins bearing the relevant sugar, 6-phosphomannose, for instance, oligonucleotides-substituted on their 5’-end with either a fluorescent probe or a radioactive label on the one hand, and bearing a thiol function on their 3’-end, on the other hand,-were coupled onto 6-phosphomannosylated proteins via a disulfide bridge. The oligonucleotide bound to 6-phosphomannosylated serum albumin is much more efficiently internalized roughly 20 times than the free oligonucleotide. Although most of the oligonucleotides are associated with vesicular compartments, oligonucleotides after releasing from the carrier by reduction of the disulfide bridge may find their way to reach the cytosol and then lead to an increase in the efficiency of the oligonucleotides.

negre, E ; Chance, ML ; Hanboula, SY ; Monsigny, M ; Roche, AC ; Mayer, RM ; Hommel, M  (1992)

Antileishmanial drug targeting through glycosylated polymers specifically internalized by macrophage membrane lectins

Antimicrobial Agents and Chemotherapy 36 (10) 2228-2232
Antileishmanial chemotherapy is hampered by the location of the parasite within the phagolysosome of the macrophage, which restricts the bioavailability of many potentially useful antileishmanial drugs. In this study, the possibility of using antileishmanial drugs targeted to the infected macrophages by means of a chemical linkage to a neutral mannose-substituted poly-L-lysine carrier molecule was explored. The study was performed in an in vitro model with Leishmania donovani-infected murine macrophages. The antileishmanial activities of various synthetic constructs were compared with those of the free drugs and the pentavalent antimonial Pentostam, which was used as the positive control. The 50% effective dose of allopurinol riboside linked to the mannosylated poly-L-lysine was below 7.5 x 10(-6) M, while it was up to 3 x 10(-4) M for the free drug, indicating that the drug bound to the polymer was 50 times more active than the free drug. Control experiments with other constructs (e.g., allopurinol riboside linked to the mannose-free polymer) confirmed that the enhancement of activity was indeed achieved by means of the mannose homing device.

Antileishmanial chemotherapy is hampered by the location of the parasite within the phagolysosome of the macrophage, which restricts the bioavailability of many potentially useful antileishmanial drugs. In this study, the possibility of using antileishmanial drugs targeted to the infected macrophages by means of a chemical linkage to a neutral mannose-substituted poly-L-lysine carrier molecule was explored. The study was performed in an in vitro model with Leishmania donovani-infected murine macrophages. The antileishmanial activities of various synthetic constructs were compared with those of the free drugs and the pentavalent antimonial Pentostam, which was used as the positive control. The 50% effective dose of allopurinol riboside linked to the mannosylated poly-L-lysine was below 7.5 x 10(-6) M, while it was up to 3 x 10(-4) M for the free drug, indicating that the drug bound to the polymer was 50 times more active than the free drug. Control experiments with other constructs (e.g., allopurinol riboside linked to the mannose-free polymer) confirmed that the enhancement of activity was indeed achieved by means of the mannose homing device.

Midoux, P ; Martin, A ; Collet, B ; Monsigny, M ; Roche, AC ; Toujas, L  (1992)

Activation of mouse macrophages by muramyl dipeptide coupled with an antimacrophage monoclonal-antibody

Bioconjugate Chemistry 3 (2) 194-199
A rat IgG2a monoclonal antibody (mAb3A33) directed against the mouse Mac-1 antigen was conjugated with muramyl dipeptide (MDP) by using an intermediate polymer ; under such conditions 75 MDP molecules were bound to one antibody molecule. A poly(L-lysine) polymer substituted with muramyl dipeptide and 3-(2-pyridyldithio)propionyl residues was prepared, the remaining lysine epsilon-amino groups were acylated with D-gluconolactone, leading to a neutral polymer ; then a few polymer conjugates were coupled to mAb3A33 via a disulfide bridge. The binding capacity of the monoclonal antibody was preserved after conjugation with MDP-polymer molecules.

A rat IgG2a monoclonal antibody (mAb3A33) directed against the mouse Mac-1 antigen was conjugated with muramyl dipeptide (MDP) by using an intermediate polymer ; under such conditions 75 MDP molecules were bound to one antibody molecule. A poly(L-lysine) polymer substituted with muramyl dipeptide and 3-(2-pyridyldithio)propionyl residues was prepared, the remaining lysine epsilon-amino groups were acylated with D-gluconolactone, leading to a neutral polymer ; then a few polymer conjugates were coupled to mAb3A33 via a disulfide bridge. The binding capacity of the monoclonal antibody was preserved after conjugation with MDP-polymer molecules.

Bonfils, E ; Mendes, C ; Roche, AC ; Monsigny, M ; Midoux, P  (1992)

Uptake by macrophages of a biotinylated oligo-a-deoxythymidylate by using mannosylated streptavidin

Bioconjugate Chemistry 3 (4) 277-284
Streptavidin substituted with mannose residues increased by 20-fold the intracellular concentration of a biotinylated dodecakis(a-deoxythymidylate) in macrophages by comparison with the uptake of free oligodeoxynucleotide. Streptavidin, the bacterial homologue of the very basic avidin, which does not contain any carbohydrate moieties and is a neutral protein, was substituted with 12 mannose residues in order to be recognized and internalized by mannose-specific lectins on the surface of macrophages. A 3'-biotinylated and 5'-fluoresceinylated dodecakis(a-deoxythymidylate) was synthesized and bound onto mannosylated streptavidin.

Streptavidin substituted with mannose residues increased by 20-fold the intracellular concentration of a biotinylated dodecakis(a-deoxythymidylate) in macrophages by comparison with the uptake of free oligodeoxynucleotide. Streptavidin, the bacterial homologue of the very basic avidin, which does not contain any carbohydrate moieties and is a neutral protein, was substituted with 12 mannose residues in order to be recognized and internalized by mannose-specific lectins on the surface of macrophages. A 3’-biotinylated and 5’-fluoresceinylated dodecakis(a-deoxythymidylate) was synthesized and bound onto mannosylated streptavidin.

Schroder, HC ; Facy, P ; Monsigny, M ; Pfeifer, K ; Bek, A ; Muller, WEG  (1992)

Purification of a glucose-binding protein from rat-liver nuclei - evidence for a role in targeting of nuclear messenger RNP to nuclear-pore complex

European Journal of Biochemistry 205 (3) 1017-1025
A nuclear carbohydrate-binding protein with a molecular mass of 67 kDa (CBP67), which is specific for glucose residues, was purified to essential homogeneity from rat liver nuclear extracts. This protein could also be isolated from nuclear ribonucleoprotein (RNP) complexes by extraction in the presence of 0.6 M or 2 M NaCl, but it was absent in polysomal RNP complex. The binding of the purified protein, which has an isoelectric point of 7.3, to glucose-containing glycoconjugates depends on the presence of Ca2+ and Mg2+.

A nuclear carbohydrate-binding protein with a molecular mass of 67 kDa (CBP67), which is specific for glucose residues, was purified to essential homogeneity from rat liver nuclear extracts. This protein could also be isolated from nuclear ribonucleoprotein (RNP) complexes by extraction in the presence of 0.6 M or 2 M NaCl, but it was absent in polysomal RNP complex. The binding of the purified protein, which has an isoelectric point of 7.3, to glucose-containing glycoconjugates depends on the presence of Ca2+ and Mg2+.

Papet, MP ; Delay, D ; Monsigny, M ; Delmotte, F  (1992)

Characterization of 2 galactosidases extracted from wheat-germ with a hydroalcoholic solvent

Biochimie 74 (1) 53-56
a- and ß-D-galactosidases were characterized from a hydroalcoholic extract of wheat germ (Triticum vulgare). Kinetic constants (V(max) and K(M)) and the optimal pHs for the hydrolysis of p-nitrophenyl galactopyranosides by both enzymes were determined. These enzymes presented a high stability in hydroalcoholic medium and were inhibited by iodoacetamide and sodium p-hydroxymercuribenzoate.

a- and ß-D-galactosidases were characterized from a hydroalcoholic extract of wheat germ (Triticum vulgare). Kinetic constants (V(max) and K(M)) and the optimal pHs for the hydrolysis of p-nitrophenyl galactopyranosides by both enzymes were determined. These enzymes presented a high stability in hydroalcoholic medium and were inhibited by iodoacetamide and sodium p-hydroxymercuribenzoate.

Cerdan, D ; Redziniak, G ; Bourgeois, CA ; Monsigny, M ; Kieda, C  (1992)

C32 human-melanoma cell endogenous lectins - characterization and implication in vesicle-mediated melanin transfer to keratinocytes

Experimental Cell Research 203 (1) 164-173


1991   Références trouvées : 10

Breton, P ; Midoux, P ; Petit, C ; Bousser, MT ; Roche, AC ; Mayer, R ; Monsigny, M  (1991)

Production of macrophage-derived cytotoxic factor by n-[3-[(carbamoylmethyl)thio]propionylated] neoglycoproteins

Bioconjugate Chemistry 2 (1) 16-18
6-Phosphomannosylated bovine serum albumin (Man6P-BSA), a neoglycoprotein endocytosed by macrophages, bearing either 3-(2-pyridyldithio)propionyl or 3-[(carbamoylmethyl)thio]propionyl residues coming from alkylation of thiol residues by iodoacetamide were prepared and tested for their immunomodulator properties. The supernatants of mouse peritoneal macrophages incubated with Man6P-BSA bearing 3-[(carbamoylmethyl)thio]propionyl groups, and by a lesser extent 3-(2-pyridyldithio)propionyl groups, were cytotoxic to L929 cells, suggesting the presence of a tumor necrosis factor like compound. This macrophage-activation process is linked to the capacity of Man6P-BSA to be endocytosed via membrane lectins of macrophages, because the supernatants of macrophages incubated with unglycosylated conjugates were not cytotoxic. The cytotoxic activity induced by 3-[(carbamoylmethyl)thio]propionyl groups bound onto Man6P-BSA was similar to that induced by Man6P-BSA bearing muramyl dipeptide, indicating that endocytosed neoglycoproteins bearing 3-](carbamoylmethyl)thio]propionyl residues are potent macrophage activators.

6-Phosphomannosylated bovine serum albumin (Man6P-BSA), a neoglycoprotein endocytosed by macrophages, bearing either 3-(2-pyridyldithio)propionyl or 3-[(carbamoylmethyl)thio]propionyl residues coming from alkylation of thiol residues by iodoacetamide were prepared and tested for their immunomodulator properties. The supernatants of mouse peritoneal macrophages incubated with Man6P-BSA bearing 3-[(carbamoylmethyl)thio]propionyl groups, and by a lesser extent 3-(2-pyridyldithio)propionyl groups, were cytotoxic to L929 cells, suggesting the presence of a tumor necrosis factor like compound. This macrophage-activation process is linked to the capacity of Man6P-BSA to be endocytosed via membrane lectins of macrophages, because the supernatants of macrophages incubated with unglycosylated conjugates were not cytotoxic. The cytotoxic activity induced by 3-[(carbamoylmethyl)thio]propionyl groups bound onto Man6P-BSA was similar to that induced by Man6P-BSA bearing muramyl dipeptide, indicating that endocytosed neoglycoproteins bearing 3-](carbamoylmethyl)thio]propionyl residues are potent macrophage activators.

Pimpaneau, V ; Midoux, P ; Monsigny, M ; Roche, AC  (1991)

Characterization and isolation of an intracellular d-mannose-specific receptor from human promyelocytic-hl-60 cells

Carbohydrate Research 213 95-108
Most mammalian macrophages express D-mannose-specific receptor (membrane lectin, M(r) 175000) allowing endocytosis of their ligands, but cells of the monocytic lineage (HL60, U937, monocyte) lack this receptor. However, after permeabilization, promyelocytic, promonocytic cells and monocytes bound fluoresceinylated D-mannose-terminated neoglycoproteins as evidenced by flow cytometry. Under these conditions, confocal analysis confirmed the intracellular membrane localization of the labeling and the absence of nuclear binding. An intracellular D-mannose-specific receptor was isolated from the human promyelocytic cell line HL60, by affinity chromatography on 4-isothiocyanatophenyl a-D-Mannopyranoside-substituted Affi-gel as a 60 000-M(r) membrane protein requiring divalent cations for the ligand binding. Under the same conditions, mouse macrophages were shown to express a 175 000-M(r) D-Mannose-specific receptor but not the 60 000-M(r) receptor.

Most mammalian macrophages express D-mannose-specific receptor (membrane lectin, M(r) 175000) allowing endocytosis of their ligands, but cells of the monocytic lineage (HL60, U937, monocyte) lack this receptor. However, after permeabilization, promyelocytic, promonocytic cells and monocytes bound fluoresceinylated D-mannose-terminated neoglycoproteins as evidenced by flow cytometry. Under these conditions, confocal analysis confirmed the intracellular membrane localization of the labeling and the absence of nuclear binding. An intracellular D-mannose-specific receptor was isolated from the human promyelocytic cell line HL60, by affinity chromatography on 4-isothiocyanatophenyl a-D-Mannopyranoside-substituted Affi-gel as a 60 000-M(r) membrane protein requiring divalent cations for the ligand binding. Under the same conditions, mouse macrophages were shown to express a 175 000-M(r) D-Mannose-specific receptor but not the 60 000-M(r) receptor.

Anjuere, F ; Monsigny, M ; Mayer, R  (1991)

Water-soluble macromolecular fluorogenic substrates for assaying proteinases - determination of pancreatic elastase activity

Analytical Biochemistry 198 (2) 342-346

Grillon, C ; Monsigny, M ; Kieda, C  (1991)

Soluble human lymphocyte sugar binding-proteins with immunosuppressive activity

Immunology Letters 28 (1) 47-56
Suppression of the immune response, which involves suppressor factors released from specialized T cells, is inhibited by a-L-rhamnose. In this paper, we show the presence of rhamnose-specific receptors on a human CD8+ T cell-rich population and describe a novel method to isolate cells which express a given sugar-binding protein on their surface. We describe the isolation of a-L-rhamnose-specific molecules (rhamnose-binding fractions : RBF) from a water-soluble extract from lymphocytes, their purification by affinity chromatography on immobilized neoglycoproteins containing rhamnose residues. RBF kept their ability to bind rhamnose, as shown by the binding of fluorescein-labeled RBF to rhamnosylated BSA-substituted beads. RBF efficiently suppresses DNA synthesis of mitogen-stimulated human lymphocytes as well as B cell immunoglobulin production. Therefore, these rhamnose-binding molecules appear to be antigen-independent suppressor factors.

Suppression of the immune response, which involves suppressor factors released from specialized T cells, is inhibited by a-L-rhamnose. In this paper, we show the presence of rhamnose-specific receptors on a human CD8+ T cell-rich population and describe a novel method to isolate cells which express a given sugar-binding protein on their surface. We describe the isolation of a-L-rhamnose-specific molecules (rhamnose-binding fractions : RBF) from a water-soluble extract from lymphocytes, their purification by affinity chromatography on immobilized neoglycoproteins containing rhamnose residues. RBF kept their ability to bind rhamnose, as shown by the binding of fluorescein-labeled RBF to rhamnosylated BSA-substituted beads. RBF efficiently suppresses DNA synthesis of mitogen-stimulated human lymphocytes as well as B cell immunoglobulin production. Therefore, these rhamnose-binding molecules appear to be antigen-independent suppressor factors.

Schrevel, J ; Barrault, C ; Grellier, P ; Mayer, R ; Monsigny, M ; Coombs, GH ; North, MJ  (1991)

Plasmodium proteinases during the erythrocytic phase of infection.

Biochemical Protozoology 270-280

Mayer, R ; Picard, I ; Lawton, P ; Grellier, P ; Barrault, C ; Monsigny, M ; Schrevel, J  (1991)

Peptide derivatives specific for a plasmodium-falciparum proteinase inhibit the human erythrocyte invasion by merozoites

Journal of Medicinal Chemistry 34 (10) 3029-3035
A specific proteinase of P. falciparum merozoites has been detected by using hydrosoluble fluorogenic peptidic substrates synthesized by classical peptide chemistry ; their N-terminal end was acylated by a gluconoyl group that protects them from aminopeptidase degradation and increases their hydrosolubility, and their carboxylic end was substituted by a 3-amino-9-ethylcarbazole group. The sequence Val-Leu-Gly-Lys was found to be the most specific substrate. On this basis, reversible peptidic inhibitors were synthesized by substituting the C-terminal lysyl residue, at the proteolytic site, by different alkylamines and amino alcohols. The activity of these compounds, studied on the P. falciparum proteinase and in in vitro cultures, strongly suggests a specific effect of this peptidic sequence on the reinvasion process. The peptidic inhibitors do not impair the release of merozoites from schizonts, but selectively inhibit the invasion step leading to the formation of rings. Although the natural target of this enzyme is not yet known, these specific peptide inhibitors could lead to a new antimalarial approach.

A specific proteinase of P. falciparum merozoites has been detected by using hydrosoluble fluorogenic peptidic substrates synthesized by classical peptide chemistry ; their N-terminal end was acylated by a gluconoyl group that protects them from aminopeptidase degradation and increases their hydrosolubility, and their carboxylic end was substituted by a 3-amino-9-ethylcarbazole group. The sequence Val-Leu-Gly-Lys was found to be the most specific substrate. On this basis, reversible peptidic inhibitors were synthesized by substituting the C-terminal lysyl residue, at the proteolytic site, by different alkylamines and amino alcohols. The activity of these compounds, studied on the P. falciparum proteinase and in in vitro cultures, strongly suggests a specific effect of this peptidic sequence on the reinvasion process. The peptidic inhibitors do not impair the release of merozoites from schizonts, but selectively inhibit the invasion step leading to the formation of rings. Although the natural target of this enzyme is not yet known, these specific peptide inhibitors could lead to a new antimalarial approach.

Harnois-Pontoni, M ; Monsigny, M ; Mayer, R  (1991)

Hydrosoluble fluorogenic substrates for plasmin

Analytical Biochemistry 193 (2) 248-255

Cerdan, D ; Grillon, C ; Monsigny, M ; Redziniak, G ; Kieda, C  (1991)

Human keratinocyte membrane lectins - characterization and modulation of their expression by cytokines

Biology of the Cell 73 (1) 35-42
In an attempt to identify cell surface molecules involved in recognition phenomena between cells such as keratinocytes and melanocytes and putatively target biological responses modifiers to keratinocytes, we undertook the detection of cell surface sugar specific receptors : membrane lectins. Keratinocyte membrane lectins were found to bind synthetic glycoproteins (neoglycoproteins) carrying either a-L-fucosyl or a-L-rhamnosyl residues. Fluorescence microscopy observations indicate that cultured keratinocytes are able to bind these two neoglycoproteins while frozen sections of human skin labelled with neoglycoprotein-coated covaspheres show that the selectivity of the binding to keratinocytes is restricted to a-L-rhamnosyl-BSA.

In an attempt to identify cell surface molecules involved in recognition phenomena between cells such as keratinocytes and melanocytes and putatively target biological responses modifiers to keratinocytes, we undertook the detection of cell surface sugar specific receptors : membrane lectins. Keratinocyte membrane lectins were found to bind synthetic glycoproteins (neoglycoproteins) carrying either a-L-fucosyl or a-L-rhamnosyl residues. Fluorescence microscopy observations indicate that cultured keratinocytes are able to bind these two neoglycoproteins while frozen sections of human skin labelled with neoglycoprotein-coated covaspheres show that the selectivity of the binding to keratinocytes is restricted to a-L-rhamnosyl-BSA.

Depierreux, C ; Kang, HC ; Guerin, B ;Monsigny, M ; Delmotte, F  (1991)

Characterization of an agrobacterium-tumefaciens lectin

Glycobiology 1 (6) 643-649
An Agrobacterium tumefaciens suspension induces a strong agglutination of aldehyde-fixed pig erythrocytes at pH 5.0. The agglutination is inhibited by some polysaccharides, such as fucoidin, and also when the pH is raised to 7.0. Lectins (sugar-binding proteins) associated with the bacterial cell wall of A. tumefaciens strain 84.5 were directly evidenced by spectrofluorimetry using fluoresceinylated neoglycoproteins. The specific binding of the fluorescein-labelled neoglycoprotein bearing .a.-L-fucoside residues was also optimal at pH 5.0. A lectin was purified by affinity chromatography on agarose substituted with .a.-L-fucopyranoside. Furthermore, the haemagglutination activity of this lectin was inhibited by polysaccharides isolated from poplar leaves.

An Agrobacterium tumefaciens suspension induces a strong agglutination of aldehyde-fixed pig erythrocytes at pH 5.0. The agglutination is inhibited by some polysaccharides, such as fucoidin, and also when the pH is raised to 7.0. Lectins (sugar-binding proteins) associated with the bacterial cell wall of A. tumefaciens strain 84.5 were directly evidenced by spectrofluorimetry using fluoresceinylated neoglycoproteins. The specific binding of the fluorescein-labelled neoglycoprotein bearing .a.-L-fucoside residues was also optimal at pH 5.0. A lectin was purified by affinity chromatography on agarose substituted with .a.-L-fucopyranoside. Furthermore, the haemagglutination activity of this lectin was inhibited by polysaccharides isolated from poplar leaves.

Grillon, C ; Monsigny, M ; Kieda, C  (1991)

Changes in the expression of lectins in human lymphocyte-T membrane upon mitogenic stimulation

Carbohydrate Research 213 283-292
Surface lectins, specific for given sugar structures, are expressed on human T cells, as shown by flow cytofluorometry using F-neoglycoproteins bearing either ß- and a-D-galactosyl, ß-D-galactosyl 6-phosphate, or a-L-rhamnosyl groups, but not by F-neoglycoproteins bearing other sugar groups (such as a-D-mannosyl groups). After stimulation with Phaseolus vulgaris mitogen, the number of cells that bind ß-D-galactosyl 6-phosphate groups (6-P-ß-D-Galp lec+ cells) increased fourfold during the first five days ; these cells are helper (CD4+) T cells. Conversely, cells that bind a-L-Rha groups belong to the T suppressor (CD8+) family and their number moderately increased. Upon stimulation by concanavalin A, the number of cells expressing the lectin recognizing a-L-Rha groups increased during the first two days and then decreased within the next two days. These results are discussed with regard to the implication of lymphocyte membrane lectins in the suppressor mechanism and in the homing process.

Surface lectins, specific for given sugar structures, are expressed on human T cells, as shown by flow cytofluorometry using F-neoglycoproteins bearing either ß- and a-D-galactosyl, ß-D-galactosyl 6-phosphate, or a-L-rhamnosyl groups, but not by F-neoglycoproteins bearing other sugar groups (such as a-D-mannosyl groups). After stimulation with Phaseolus vulgaris mitogen, the number of cells that bind ß-D-galactosyl 6-phosphate groups (6-P-ß-D-Galp lec+ cells) increased fourfold during the first five days ; these cells are helper (CD4+) T cells. Conversely, cells that bind a-L-Rha groups belong to the T suppressor (CD8+) family and their number moderately increased. Upon stimulation by concanavalin A, the number of cells expressing the lectin recognizing a-L-Rha groups increased during the first two days and then decreased within the next two days. These results are discussed with regard to the implication of lymphocyte membrane lectins in the suppressor mechanism and in the homing process.


1990   Références trouvées : 13

Petit, C ; Monsigny, M ; Roche, AC  (1990)

Macrophage activation by muramyl dipeptide bound to neoglycoproteins and glycosylated polymers - cytotoxic factor production

Journal of Biological Response Modifiers 9 (1) 33-43

Roche, AC ; Midoux, P ; Pimpaneau, V ; Negre, E ; Mayer, R ; Monsigny, M  (1990)

Endocytosis mediated by monocyte and macrophage membrane lectins - application to antiviral drug targeting

Research in Virology 141 (2) 243-249

Breton, P ; Monsigny, M ; Mayer, R  (1990)

Synthesis of new acylpseudopeptides analogous to Nn-acetylmuramyl dipeptide (MDP)

Tetrahedron 46 (12) 4265-4276

Breton, P ; Monsigny, M ; Mayer, R  (1990)

PSI [CH2O] pseudodipeptide synthesis - an improved approach which allows absolute-configuration determination

International Journal of Peptide and Protein Research 35 (4) 346-351

Schrevel, J ; Deguercy, A ; Mayer, R ; Monsigny, M  (1990)

Proteases in malaria-infected red-blood-cells

Blood Cells 16 (2-3) 563-584

Kieda, C ; Monsigny, M  (1990)

Lymphocyte membrane lectins - their role in lymphocyte homing

Eos-Rivista Di Immunologia Ed Immunofarmacologia 10 (4) 172-172

Midoux, P ; Negre, E ; Roche, AC ; Mayer, R ; Monsigny, M ; Balzarini, J ; Declercq, E ; Mayer, E ; Ghaffar, A ; Gangemi, JD  (1990)

Drug targeting - anti-HDV-1 activity of mannosylated polymer-bound 9-(2-phosphonylmethoxyethyl)adenine

Biochemical and Biophysical Research Communications 167 (3) 1044-1049

Bourgeois, CA ; Monsigny, M  (1990)

Distribution of sugar-binding sites within interphase nuclei and mitotic chromosomes of a human cell-line

Biology of The Cell 69 (2) 119-126

Papet, MP ; Fournet, B ; Monsigny, M ; Delmotte, F  (1990)

Characterization of a high molecular mass cytokinin-like compound extracted from wheat-germ (triticum-vulgare L)

Plant Science 68 (2) 175-182

Grillon C ; Monsigny M ; Kieda C  (1990)

Cell surface lectins of human granulocytes their expression is modulated by mononuclear cells and granulocyte-macrophage colony-stimulating factor

Glycobiology 1 (1) 33-38
This paper presents the characterization of a sugar-specific receptor on the surface of human circulating polymorphonuclear cells. With the help of fluorescent neoglycoproteins and flow cytometry, a receptor was identified as being specific for .a.-L-rhamnosyl residues. The number of receptors was 55 000/cell and their affinity reached 2 .times. 108 1 mol-1. This number changes as a function of the biological state of the cells. Indeed, receptor expression was modulated by the presence of other cells. T cells and B cells increased the number of receptors on the granulocyte surface. Expression of the .a.-L-rhamnose-specific lectin was dependent on lymphocyte derived soluble factor(s), which induce(s) growth and differentiation of polymorphonuclear phagocytes. Granulocyte/macrophage colony-stimulating factor (GM-CSF) specifically produced a significant increase in the number of receptors for .a.-L-rhamnose (2-10-fold/cell). This modulation was independent of protein kinase C activators such as phorbol ester, which produced no effect on .a.-L-rhamnose receptor expression. These findings demonstrate that GM-CSF may stimulate post differentiation functions and properties of mature granulocytes.

This paper presents the characterization of a sugar-specific receptor on the surface of human circulating polymorphonuclear cells. With the help of fluorescent neoglycoproteins and flow cytometry, a receptor was identified as being specific for .a.-L-rhamnosyl residues. The number of receptors was 55 000/cell and their affinity reached 2 .times. 108 1 mol-1. This number changes as a function of the biological state of the cells. Indeed, receptor expression was modulated by the presence of other cells. T cells and B cells increased the number of receptors on the granulocyte surface. Expression of the .a.-L-rhamnose-specific lectin was dependent on lymphocyte derived soluble factor(s), which induce(s) growth and differentiation of polymorphonuclear phagocytes. Granulocyte/macrophage colony-stimulating factor (GM-CSF) specifically produced a significant increase in the number of receptors for .a.-L-rhamnose (2-10-fold/cell). This modulation was independent of protein kinase C activators such as phorbol ester, which produced no effect on .a.-L-rhamnose receptor expression. These findings demonstrate that GM-CSF may stimulate post differentiation functions and properties of mature granulocytes.

Depierreux, C ; Lebris, MT ; Michel, MF ; Valeur, B ; Monsigny, M ; Delmotte, F  (1990)

Benzoxazinone kanamycin derivative - a new fluorescent-probe for flow-cytometry analysis of bacteria (agrobacterium-tumefaciens)

Fems Microbiology Letters 67 (3) 237-244

Monsigny, M ; Midoux, P ; Depierreux, C ; Bebear, C ; Lebris, MT ; Valeur, B  (1990)

Benzoxazinone kanamycin-a conjugate - a new fluorescent-probe suitable to detect mycoplasmas in cell-culture

Biology of The Cell 70 (3) 101-105
The synthesis of a new benzoxazinone derivative suitable to detect early infection of cultured cells with mycoplasmas is described. p-[ß-(7-dimethylamino 1,4-benzoxazin 2-one 3yl)-vinyl]-phenylpropenoic acid was coupled to kanamycin A, an aminoglycoside leading to a cationic fluorescent probe which fluoresces at 600 nm upon excitation at 490 nm. This fluorescent probe is shown to heavily label the glycocallix of all the mycoplasma strains tested which are found to be associated with contaminated cultured cells and to allow an easy and rapid detection of contamination by fluorescence microscopy and flow cytometry.

The synthesis of a new benzoxazinone derivative suitable to detect early infection of cultured cells with mycoplasmas is described. p-[ß-(7-dimethylamino 1,4-benzoxazin 2-one 3yl)-vinyl]-phenylpropenoic acid was coupled to kanamycin A, an aminoglycoside leading to a cationic fluorescent probe which fluoresces at 600 nm upon excitation at 490 nm. This fluorescent probe is shown to heavily label the glycocallix of all the mycoplasma strains tested which are found to be associated with contaminated cultured cells and to allow an easy and rapid detection of contamination by fluorescence microscopy and flow cytometry.

Facy, P ; Seve, AP ; Hubert, M ; Monsigny, M ; Hubert, J  (1990)

Analysis of nuclear sugar-binding components in undifferentiated and in vitro differentiated human promyelocytic leukemia-cells (hl60)

Experimental Cell Research 190 (2) 151-160


1989   Références trouvées : 6

Derrien, D ; Midoux, P ; Petit, C ; Negre, E ; Mayer, R ; Monsigny, M ; Roche, AC  (1989)

Muramyl dipeptide bound to poly-l-lysine substituted with mannose and gluconoyl residues as macrophage activators

Glycoconjugate Journal 6 (2) 241-255

Midoux, P ; Petit, C ; Pellen, P ; Toujas, L ; Monsigny, M ; Roche, AC  (1989)

Macrophage antigens associated with adhesion - identification by a monoclonal-antibody specific for lewis lung-carcinoma cells

Experimental Cell Research 183 (1) 168-178

Monsigny, M ; Midoux, P ; Lebris, MT ; Roche, AC ; Valeur, B  (1989)

Benzoxazinone derivatives - new fluorescent-probes for 2-color flow-cytometry analysis using ole excitation wavelength

Biology of The Cell 67 (2) 193-200

Grellier, P ; Picard, I ; Bernard, F ; Mayer, R ; Heidrich, HG ; Monsigny, M ; Schrevel, J  (1989)

Purification and indentification of a neutral endopeptidase in plasmodium-falciparum schizonts and merozoites

Parasitology Research 75 (6) 455-460

Pimpaneau, V ; Midoux, P ; Durand, G ; Debaetselier, P ; Monsigny, M ; Roche, AC  (1989)

Endocytosis o f a-1-acid glycoprotein variants and of neoglycoproteins containing mannose derivatives by a mouse hybridoma cell-line (2c11-12) - comparison with mouse peritoneal-macrophages

Glycoconjugate Journal 6 (4) 561-574

Hubert, J ; Seve, AP ; Facy, P ; Monsigny, M  (1989)

Are nuclear lectins and nuclear glycoproteins involved in the modulation of nuclear functions ?

Cell Differentiation and Development 27 (2) 69-81


1988   Références trouvées : 9

Monsigny, M ; Petit, C ; Roche, AC  (1988)

Colorimetric determination of neutral sugars by a resorcinol sulfuric-acid micromethod

Analytical Biochemistry 175 (2) 525-530

Monsigny, M ; Roche, AC ; Kieda, C ; Midoux, P ; Obrenovitch, A  (1988)

Characterization and biological implications of membrane lectins in tumor, lymphoid and myeloid cells

Biochimie 70 (11) 1633-1649

Latge, JP ; Monsigny, M ; Prevost, MC  (1988)

Visualization of exocellular lectins in the entomopathogenic fungus conidiobolus-obscurus

Journal of Histochemistry & Cytochemistry 36 (11) 1419-1424

Daussin, F ; Boschetti, E ; Delmotte, F ; Monsigny, M  (1988)

Para-benzylthiocarbamoyl-aspartyl-daunorubicin-substituted polytrisacryl - a new drug acid-labile arm-carrier conjugate

European Journal of Biochemistry 176 (3) 625-628

Schrevel, J ; Grellier, P ; Mayer, R ; Monsigny, M  (1988)

Neutral proteases involved in the reinvasion of erythrocytes by plasmodium merozoites

Biology of The Cell 64 (2) 233-244

Olins, DE ; Olins, AL ; Seve, AP ; Bourgeois, CA ; Hubert, J ; Monsigny, M  (1988)

Lectin-like components in the macronuclear replication bands of euplotes-eurystomus

Biology of The Cell 62 (1) 95-98

Almahmood, S ; Giumelly, P ; Bonaly, R ; Delmotte, F ; Monsigny, M  (1988)

Kluyveromyces-bulgaricus yeast lectins - isolation of N-acetylglucosamine and galactose-specific lectins - their relation with flocculation

Journal of Biological Chemistry 263 (8) 3930-3934

Alquier, C ; Miquelis, R ; Monsigny, M ; Athouelhaon, AM  (1988)

Direct fluorescence localization of an endogenous N-acetyl-glucosamine-specific lectin in the thyroid-gland

Histochemistry 89 (2) 171-176

Monsigny, M ; Petit, C ; Roche, AC  (1988)

Detection of lectin-binding sites and endogenous lectins by glycosylated affinity markers

Analytical Biochemistry 175 (2) 525-530


1987   Références trouvées : 7

Midoux, P ; Roche, AC ; Monsigny, M  (1987)

Quantitation of the binding, uptake, and degradation of fluoresceinylated neoglycoproteins by flow-cytometry

Cytometry 8 (3) 327-334

Miquelis, R ; Alquier, C ; Monsigny, M  (1987)

The N-acetylglucosamine-specific receptor of the thyroid - binding characteristics, partial characterization, and potential role

Journal of Biological Chemistry 262 (31) 15291-15298

Bernard, F ; Mayer, R ; Picard, I ; Deguercy, A ; Monsigny, M ; Schrevel, J  (1987)

Plasmodium-berghei and plasmodium-chabaudi - a neutral endopeptidase in parasite extracts and plasma of infected animals

Experimental Parasitology 64 (1) 95-103

Bourgeois, C ; Seve, AP ; Monsigny, M ; Hubert, J  (1987)

Localization of lectins within the nucleolus and the interchromatin spaces in mammalian-cell nuclei

Biology of The Cell 60 (1) A28-A28

Bourgeois, CA ; Seve, AP ; Monsigny, M ; Hubert, J  (1987)

Detection of sugar-binding sites in the fibrillar and the granular components of the nucleolus an experimental study in cultured mammalian cells

Experimental Cell Research 172 (2) 365-376
The intranucleolar distribution of sugar-binding sites (i.e., lectin-like molecules) was analyzed in segregated nucleoli of actinomycin D-treated HeLa cells. The detection of sugar-binding sites was performed by incubation either of permeabilized nuclei in the presence of fluorescein-labeled neoglycoproteins or of ultrathin sections cut through in situ-fixed nuclei in the presence of gold-labeled neoglycoproteins. In the former case, the fluorescent nucleolar components were identified by comparison with the nucleolar components of similarly treated cells observed in electron microscopy. For the first time, this study reveals the presence of sugar-binding sites in both the fibrillar and the granular components of the nucleolus.

The intranucleolar distribution of sugar-binding sites (i.e., lectin-like molecules) was analyzed in segregated nucleoli of actinomycin D-treated HeLa cells. The detection of sugar-binding sites was performed by incubation either of permeabilized nuclei in the presence of fluorescein-labeled neoglycoproteins or of ultrathin sections cut through in situ-fixed nuclei in the presence of gold-labeled neoglycoproteins. In the former case, the fluorescent nucleolar components were identified by comparison with the nucleolar components of similarly treated cells observed in electron microscopy. For the first time, this study reveals the presence of sugar-binding sites in both the fibrillar and the granular components of the nucleolus.

Gabius, HJ ; Engelhardt, R ; Hellmann, T ; Midoux, P ; Monsigny, M ; Nagel, GA ; Vehmeyer, K  (1987)

Characterization of membrane lectins in human-colon carcinoma-cells by flow cytofluorometry, drug targeting and affinity-chromatography

Anticancer Research 7 (1) 109-112

Singh, B ; Monsigny, M ; Hommel, M  (1987)

Amino-sugars inhibit the in vitro cytoadherence of plasmodium-falciparum-infected erythrocytes to melanoma-cells

Molecular and Biochemical Parasitology 23 (1) 47-53


1986   Références trouvées : 18

Seve, A.P., Hubert, J., Bouvier, D., Bourgeois, C., Midoux, P., Roche, A.C. & Monsigny, M.  (1986)

Analysis of sugar-binding sites in mammalian cell nuclei by quantitative flow microfluorometry

Proc. Natl. Acad. Sci. USA 83, 5997-6001

Delmotte, F ; Mayer, R ; Lescanne, PJ ; Pontoni, M ; Roche, AC ; Monsigny, M  (1986)

Micromolecular and macromolecular hydrosoluble prodrugs containing adriamycin and daunomycin

Cancer Drug Delivery 3 (1) 82-82

Roche, AC ; Monsigny, M  (1986)

Macrophage activation by targeted biological response modifiers

Annales de L Institut Pasteur-Immunology C137 (2) 223-226

Mayer, R ; Pontoni, M ; Roche, AC ; Monsigny, M  (1986)

In vitro cytotoxicity of antitumoral prodrugs

Cancer Drug Delivery 3 (1) 83-83

Roche, AC ; Midoux, P ; Petit, C ; Mayer, R ; Monsigny, M ; Patchen, ML  (1986)

Immunostimulating drugs targeted by neoglycoproteins - effects on hematopoiesis and therapeutic properties

Bulletin Du Cancer 73 (4) 424-424

Lepoivre, M ; Roche, AC ; Tenu, JP ; Petit, JF ; Nolibe, D ; Monsigny, M  (1986)

Identification of 2 macrophage populations by flow-cytometry monitoring of oxidative burst and phagocytic functions

Biology of The Cell 57 (2) 143-146

Midoux, P ; Roche, AC ; Monsigny, M  (1986)

Estimation of the degradation of endocytosed material by flow cytofluorometry using 2 neoglycoproteins containing different numbers of fluorescein molecules

Biology of The Cell 58 (3) 221-225

Obrenovitch, A ; Monsigny, M  (1986)

Tumor angiogenesis

Pathologie Biologie 34 (3) 189-201

Boschetti, E ; Egly, JM ; Monsigny, M  (1986)

The place of affinity-chromatography in the production and purification of biomolecules from cultured-cells

Trac-Trends in Analytical Chemistry 5 (1) 4-10

Schrevel, J ; Philippe, M ; Bernard, F ; Monsigny, M  (1986)

Surface plasmodium sugar-binding components evidenced by fluorescent neoglycoproteins

Biology of The Cell 56 (1) 49-55

Petit, PX ; Perennes, C ; Bergounioux, C ; Seve, AP ; Delmotte, F ; Monsigny, M  (1986)

Quantitative-analysis of sugar-binding proteins in plant-cell nuclei by flow-cytometry

Biology of The Cell 58 (3) A15-A15

Delmotte, F ; Lescanne, PJ ; Daussin, F ; Monsigny, M  (1986)

Macromolecular prodrugs - antitumor-activity of peptidyl daunorubicin linked to highly hydrosoluble polymers

Cancer Drug Delivery 3 (1) 77-77

Kieda, C ; Monsigny, M  (1986)

Involvement of membrane sugar receptors and membrane glycoconjugates in the adhesion of 3LL-cell subpopulations to cultured pulmonary cells

Invasion & Metastasis 6 (6) 347-366

Kieda, CM ; Monsigny, M  (1986)

Involvement of membrane lectins in the establishment of metastases of lewis lung-carcinoma cells

Bulletin Du Cancer 73 (2) 221-222

Kieda, C ; Monsigny, M  (1986)

Involvement of membrane lectins in cell-cell recognition - pulmonary cells and lewis lung-carcinoma cells

Bulletin Du Cancer 73 (4) 419-419

El Kebbaj, MS ; Latruffe, N ; Monsigny, M ; Obrenovitch, A  (1986)

Interactions between APO-(D-ß-hydroxybutyrate dehydrogenase) and phospholipids studied by intrinsic and extrinsic fluorescence

Biochemical Journal 237 (2) 359-364

Schroder, HC ; Becker, R ; Bachmann, M ; Gramzow, M ; Seve, AP ; Monsigny, M ; Muller, WEG  (1986)

Differential changes of nuclear-envelope-associated enzyme-activities involved in nucleocytoplasmic messenger-MA transport in the developing rat-brain and liver

Biochimica et Biophysica Acta 868 (2-3) 108-118

Miquelis, R ; Alquier, C ; Monsigny, M  (1986)

Alectin-like n-acetylglucosamine receptor on thyrocytes - binding characteristics

Annales D’endocrinologie 47 (6) 65-65


1985   Références trouvées : 8

Monsigny, M ; Midoux, P ; Roche, AC  (1985)

Use of flow-cytometry in the study of endocytosis

Biology of The Cell 55 (1-2) A23-A23

Maillet, T ; Roche, AC ; Therain, F ; Monsigny, M  (1985)

Time course localization of immunoglobulin-m monoclonal-antibody and its fragments in leukemic tumor-bearing mice

Cancer Immunology Immunotherapy 19 (3) 177-182

Roche, AC ; Monsigny, M  (1985)

Sugar binding-protein (membrane lectin) on human-monocytes

Journal of Leukocyte Biology 38 (1) 102-102

Roche, AC ; Midoux, P ; Bouchard, P ; Monsigny, M  (1985)

Membrane lectins on human-monocytes - maturation-dependent modulation of 6-phosphomannose and mannose receptors

Febs Letters 193 (1) 63-68

Roche, AC ; Bailly, P ; Monsigny, M  (1985)

Macrophage activation by mdp bound to neoglycoproteins - metastasis eradication in mice

Invasion & Metastasis 5 (4) 218-232

Roche, AC ; Mayer, R ; Midoux, P ; Monsigny, M  (1985)

Immunostimulant drugs targeted by monoclonal-antibodies and neoglycoproteins

Journal of Leukocyte Biology 38 (1) 138-138

Hubert, J ; Seve, AP ; Bouvier, D ; Masson, C ; Bouteille, M ; Monsigny, M  (1985)

In situ ultrastructural-localization of sugar-binding sites in lizard granulosa-cell nuclei

Biology of The Cell 55 (1-2) 15-20

Seve, AP ; Hubert, J ; Bouvier, D ; Bouteille, M ; Maintier, C ; Monsigny, M  (1985)

Detection of sugar-binding proteins in membrane-depleted nuclei

Experimental Cell Research 157 (2) 533-538


1984   Références trouvées : 23

Monsigny, M ; Roche, AC ; Midoux, P  (1984)

Uptake of neoglycoproteins via membrane lectin(s) of l1210 cells evidenced by quantitative flow cytofluorometry and drug targeting

Biology of The Cell 51 (2) 187-196

Midoux, P ; Maillet, T ; Therain, F ; Monsigny, M ; Roche, AC  (1984)

Tumor-localization of lewis lung-carcinoma with radiolabeled monoclonal-antibodies

Cancer Immunology Immunotherapy 18 (1) 19-23

Monsigny, M ; Roche, AC ; Bailly, P  (1984)

Tumoricidal activation of murine alveolar macrophages by muramyldipeptide substituted mannosylated serum-albumin

Biochemical and Biophysical Research Communications 121 (2) 579-584

Roche, AC ; Bailly, P ; Midoux, P ; Monsigny, M  (1984)

Selective macrophage activation by muramyldipeptide bound to monoclonal-antibodies specific for mouse-tumor cells

Cancer Immunology Immunotherapy 18 (3) 155-159

Monsigny, M., Roche, A.C. and Midoux, P.  (1984)

Monoclonal-antibodies (igm) against a murine carcinoma, lewis lung-carcinoma (3ll) - production, purification and in vitro selectivity

Immunology Letters 8 (3) 131-136

Midoux, P ; Roche, AC ; Monsigny, M  (1984)

Flow cytofluorometry analysis of neoglycoproteins endocytosis mediated by membrane lectin of lewis lung-carcinoma cells

Biology of The Cell 52 (2) A76-A76

Roche, AC ; Bailly, P ; Monsigny, M  (1984)

Enhancement of macrophages activation by membrane lectin mediated endocytosis of muramyldipeptide bound to neoglycoproteins

Biology of The Cell 51 (2) A58-A58

Roche, AC ; Midoux, P ; Lescanne, PJ ; Marteau, C ; Delmotte, F ; Monsigny, M  (1984)

Drug targeting via membrane lectins of mouse leukemic and carcinoma-cells

Biology of The Cell 51 (2) A59-A59

Latge, JP ; Monsigny, M ; Prevost, MC ; Roche, AC ; Kieda, C ; Fournet, B  (1984)

Carbohydrate-binding proteins in the entomogenous fungus conidiobolus-obscurus

Biology of The Cell 51 (2) A52-A52

Midoux, P ; Grivet, JP ; Delmotte, F ; Monsigny, M  (1984)

The binding of monosaccharides to wheat-germ-agglutinin - fluorescence and NMR investigations

Biochemical and Biophysical Research Communications 119 (2) 603-611

Kieda, C ; Monsigny, M  (1984)

Specific membrane lectin of human T-suppressor cells

Immunobiology 167 (1-3) 87-88

Monsigny, M  (1984)

Special issue - the role of carbohydrates in cell recognition - endogenous lectins - foreword

Biology of The Cell 51 (2) R4-R4

Philippe, M ; Midoux, P ; Hommel, M ; Monsigny, M ; Schrevel, J  (1984)

Preliminary-study of glycosidases during the endoerythrocytary phase of plasmodium

Journal of Protozoology 31 (4) A75-A75

Kieda, C ; Monsigny, M  (1984)

Possible involvement of membrane lectins in lung metastasis of lewis lung-carcinoma cells(3LL)

Biology of The Cell 51 (2) A51-A51

Monsigny M  (1984)

International meeting on the role of carbohydrates in cell recognition endogenous lectins aussois france march 18-24

Biology of The Cell 51 (2) 113-294 46a-62a

Midoux, P ; Wahl, P ; Auchet, JC ; Monsigny, M  (1984)

Fluorescence quenching of tryptophan by trifluoroacetamide

Biochimica et Biophysica Acta 801 (1) 16-25

Seve, AP ; Hubert, J ; Bouvier, D ; Bouteille, M ; Maintier, C ; Monsigny, M  (1984)

Evidence of nuclear endogenous lectins by means of neoglycoproteins

Biology of The Cell 52 (2) A90-A90

Seve, AP ; Hubert, J ; Bouvier, D ; Bouteille, M ; Maintier, C ; Monsigny, M  (1984)

Evidence and quantitative-analysis by means of cytometry in the flow of endogenic nuclear lectins

Biology of The Cell 52 (3) A132-A132

Bernard, F ; Monsigny, M ; Schrevel, J  (1984)

Evidence and classification of an endopeptidase with a strong affinity for the VAL-LAU-GLY-ARG-AEC fluorogenic peptide in plasmodium in rodents

Journal of Protozoology 31 (4) A64-A64

Krajhanzl, A ; Nosek, J ; Monsigny, M ; Kocourek, J  (1984)

Direct visualization of endogenous lectins in fish oocytes by glycosylated fluorescent cytochemical markers

Histochemical Journal 16 (4) 426-428

Seve, AP ; Bouvier, D ; Hubert, J ; Bouteille, M ; Monsigny, M  (1984)

Detection of nuclear endogenous lectins by neoglycoprotein binding

Biology of The Cell 51 (2) A60-A60

Bre, MH ; Leforttran, M ; Obrenovitch, A ; Monsigny, M  (1984)

Detection of euglena cell-surface carbohydrates by lectins - alterations related to vitamin-B12 deficiency

European Journal of Cell Biology 35 (2) 273-278

Schrevel, J ; Bernard, F ; Maintier, C ; Mayer, R ; Monsigny, M  (1984)

Detection and characterization of a selective endopeptidase from plasmodium-berghei by using fluorogenic peptidyl substrates

Biochemical and Biophysical Research Communications 124 (3) 703-710


1983   Références trouvées : 14

Roche, AC ; Barzilay, M ; Midoux, P ; Junqua, S ; Sharon, N ; Monsigny, M  (1983)

Sugar-specific endocytosis of glycoproteins by lewis lung-carcinoma cells

Journal of Cellular Biochemistry 22 (3) 131-140

Midoux, P ; Roche, AC ; Monsigny, M  (1983)

Preparation of monoclonal-antibodies directed against tumoral cells of a murine carcinoma, pulmonary lewis carcinoma

Biology of The Cell 49 (1) A33-A33

Monsigny, M ; Kieda, C ; Roche, AC  (1983)

Membrane-glycoproteins glycolipids and membrane lectins as recognition signals in normal and malignant-cells

Biology of The Cell 47 (1) 95-110

Maillet, T ; Therain, F ; Midoux, P ; Bailly, P ; Roche, AC ; Monsigny, M  (1983)

In vivo study of the localization of tumoral anti-cellular monoclonal-antibodies in mice

Biology of The Cell 49 (1) A32-A32

Schrevel, J ; Bernard, F ; Monsigny, M  (1983)

Targeting of peptidyl drugs in plasmodium

Journal of Protozoology 30 (3) A73-A74

Obrenovitch, A ; Maintier, C ; Maillet, T ; Mayer, R ; Kieda, C ; Monsigny, M  (1983)

Sensitive fluorometric-determination of plasminogen-activator in cell lysates and supernatants

Febs Letters 157 (2) 265-270

Bladier, D ; Vassy, R ; Perret, G ; Cornillot, P ; Monsigny, M  (1983)

Red-cell aging - phagocytosis and life-span of young and old erythrocytes fractionated by centrifugation

Biology of The Cell 49 (3) 231-235

Midoux, P ; Delmotte, F ; GRIVET, JP ; Monsigny, M  (1983)

Protein sugar interactions - environmental-effect on the fluorescence of ortho-(4-methylumbelliferyl)-glycosides

Biochemical and Biophysical Research Communications 110 (3) 926-933

Kieda, C ; Monsigny, M  (1983)

Participation of membrane lectins of lymphocytes with phenomena of specific adhesion to endothelial-cells of post-capillary venules of lymphatic ganglia

Biology of The Cell 49 (1) A31-A31

Obrenovitch A ; Maintier C ; Sene C ; Boschetti E ; Monsigny M  (1983)

Micro carrier culture of fibroblastic cells on modified tris acryl beads

Biology of The Cell 46 (3) 249-256
New microcarriers (Trisacryl beads) based on the copolymerization of 2-acrylamido-2-hydroxymethylpropane-1,3-diol and N,N'-diallyltartradiamide were synthesized in order to grow cells requiring attachment to a solid matrix. These microcarriers have a very low content of ionic charges, their density is .apprx. 1.05 and they are only permeable to small molecules ; they are devoid of cytotoxicity. Their hydrophilicity prevents binding or adsorption of cells onto their surface. Gelatin-coupled Trisacryl beads and heparin-substituted Trisacryl beads were prepared. These gelatin or heparin Trisacryl beads are sterilized by autoclaving. BHK 21 C 13 cells readily attach to and spread on gelatin-coupled Trisacryl beads and on fibronectin-precoated heparin-substituted Trisacryl beads. The growth rate of BHK 21 C 13 cells attached on gelatin-coupled Triascryl beads is as high as the growth rate of cells attached to plastic culture dishes (doubling time of 14 h). Cells attached to gelatin-coupled Trisacryl beads are detached by a trypsin-disodium EDTA treatment. When beads covered with BHK 21 C 13 cells were inoculated in culture dishes containing cell-free gelatin-coupled Trisacryl beads, cells spontaneously colonized the cell-free beads.

New microcarriers (Trisacryl beads) based on the copolymerization of 2-acrylamido-2-hydroxymethylpropane-1,3-diol and N,N’-diallyltartradiamide were synthesized in order to grow cells requiring attachment to a solid matrix. These microcarriers have a very low content of ionic charges, their density is .apprx. 1.05 and they are only permeable to small molecules ; they are devoid of cytotoxicity. Their hydrophilicity prevents binding or adsorption of cells onto their surface. Gelatin-coupled Trisacryl beads and heparin-substituted Trisacryl beads were prepared. These gelatin or heparin Trisacryl beads are sterilized by autoclaving. BHK 21 C 13 cells readily attach to and spread on gelatin-coupled Trisacryl beads and on fibronectin-precoated heparin-substituted Trisacryl beads. The growth rate of BHK 21 C 13 cells attached on gelatin-coupled Triascryl beads is as high as the growth rate of cells attached to plastic culture dishes (doubling time of 14 h). Cells attached to gelatin-coupled Trisacryl beads are detached by a trypsin-disodium EDTA treatment. When beads covered with BHK 21 C 13 cells were inoculated in culture dishes containing cell-free gelatin-coupled Trisacryl beads, cells spontaneously colonized the cell-free beads.

Obrenovitch, A ; Maintier, C ; Maillet, T ; Mayer, R ; Kieda, C ; Monsigny, M  (1983)

High-sensitivity fluorometric method for the detection of the plasminogen-activator

Biology of The Cell 49 (1) A13-A13

Gros, D ; Bruce, B ; Kieda, C ; Delmotte, F ; Monsigny, M ; Schrevel, J  (1983)

Evolution of the surface of mouse myocardial-cells during ontogenesis .2. Changes of fluorescent lectin-binding sites

Journal of Molecular and Cellular Cardiology 15 (2) 93-104

Bre, MH ; Obrenovitch, A ; Monsigny, M ; Dubuc, N ; Pouphile, M ; Leforttran, M  (1983)

Analysis of cortical glycoproteins of normal euglenas and euglenas with a vitamin-b12 deficiency using fluorescent lecithins - microscopy and flux cytometry

Biology of The Cell 49 (1) A2-A2

Roos, PH ; Kolbbachofen, V ; Schlepperschafer, J ; Monsigny, M ; Stockert, RJ ; Kolb, H  (1983)

2 galactose-specific receptors in the liver with different function

Febs Letters 157 (2) 253-256


1982   Références trouvées : 8

Roche, AC ; Monsigny, M  (1982)

Monoclonal-antibody (igm)specific to mouse lymphoma-cells (l 1210) - isolation and therapeutic properties

Cell Biology International Reports 6 (6) 557-565

Tenu, JP ; Roche, AC ; Yapo, A ; Kieda, C ; Monsigny, M ; Petit, JF  (1982)

Absence of cell-surface receptors for muramylpeptides in mouse peritoneal-macrophages

Biology of The Cell 44 (2) 157-164

Kieda, C ; Monsigny, M  (1982)

The participation of lymphocyte membrane sugar receptors (endogeneous lectins) in their adhesion to the endothelial-cells of lymph-node veinules

Immunobiology 163 (2-4) 244-244

Obrenovitch, A ; Sene, C ; Maintier, C ; Boschetti, E ; Monsigny, M  (1982)

New microcarriers suitable for the culture of fibroblastic cells

Biology of The Cell 45 28-28

Obrenovitch, A ; Maintier, C ; Sene, C ; Boschetti, E ; Monsigny, M  (1982)

Microcarrier culture of fibroblastic cells on modified trisacryl beads

Biology of The Cell 46 (3) 249-256

Monsigny, M  (1982)

Characterization and possible function of membrane lectins

Fresenius Zeitschrift Fur Analytische Chemie 311 (4) 331-331

Waxdal, MJ ; Kieda, C ; Monsigny, M  (1982)

Cell-surface carbohydrate receptors (endogeneous lectins) - a new class of differentiation markers for lymphoid-cells

Immunobiology 163 (2-4) 211-211

Monsigny, M ; Kieda, C ; Maillet, T  (1982)

Assay for proteolytic activity using a new fluorogenic substrate (peptidyl-3-amino-9-ethyl-carbazole) - quantitative-determination of lipopolysaccharide at the level of one picogram

Embo Journal 1 (3) 303-306


1981   Références trouvées : 6

Magetdana, R ; Veh, RW ; Sander, M ; Roche, AC ; Schauer, R ; Monsigny, M  (1981)

Specificities of limulin and wheat-germ-agglutinin towards some derivatives of gm3 gangliosides

European Journal of Biochemistry 114 (1) 11-16

Roche, AC ; Monsigny, M  (1981)

Preparation and properties of anti-l-1210 monoclonal-antibodies

Biology of The Cell 42 (1) A4-A4

Obrenovitch, A ; Sene, C ; Roche, AC ; Monsigny, M ; Visher, P ; Hughes, RC  (1981)

Cell-surface receptors for wheat-germ-agglutinin and limulin in baby hamster-kidney cells and ricin resistant variants

Biochimie 63 (3) 169-175

Schulte, H ; Monsigny, M  (1981)

Nuclear-membrane lectins of rat-liver - direct visualization with fluorescent glycosylated markers

Biology of The Cell 42 (1) 13-17

Courtot, AM ; Auroux, M ; Obrenovitch, A ; Monsigny, M  (1981)

Intraindividual variability of concanavalin-a binding to the plasmalemma of human-spermatozoa

Archives of Andrology 7 (4) 307-311

Schrevel, J ; Gros, D ; Monsigny, M  (1981)

Cyto-chemistry of cell glycoconjugates

Progress in Histochemistry and Cytochemistry 14 (2) 1-269


1980   Références trouvées : 4

Goldstein, IJ ; Hughes, RC ; Monsigny, M ; Osawa, T ; Sharon, N  (1980)

What should be called a lectin ?

Nature 285 (5760) 66-66

Monsigny, M ; Roche, AC ; Sene, C ; Maget-Dana, R ; Delmotte, F  (1980)

Sugar-lectin interactions - How does wheat-germ-agglutinin bind sialoglycoconjugates ?

European Journal of Biochemistry 104 (1) 147-153

Monsigny, M ; Kieda, C ; Roche, AC ; Delmotte, F  (1980)

Preparation and biological properties of a covalent anti-tumor drug-ARM-carrier (DACconjugate)

Febs Letters 119 (1) 181-186

Midoux, P ; Grivet, JP ; Monsigny, M  (1980)

Lectin-sugar interactions - the binding of 1-0-methyl-di-N-trifluoroacetyl-ß-chitobioside to wheat-germ-agglutinin

Febs Letters 120 (1) 29-32


1979   Références trouvées : 15

Roche, AC ; Delmotte, F ; magetdana, R ; Monsigny, M  (1979)

Specificities of n-acetyl neuraminic acid binding lectins - limulin and wheat-germ-agglutinin

Biologie Cellulaire 36 (3) A9-A9

Monsigny, M ; Sene, C ; Obrenovitch, A ; Roche, AC ; Delmotte, F ; Boschetti, E  (1979)

Properties of succinylated wheat-germ-agglutinin

European Journal of Biochemistry 98 (1) 39-45

Monsigny, M ; Kieda, C ; Roche, AC  (1979)

Membrane lectins

Biologie Cellulaire 36 (3) 289-300

Kieda, C ; Roche, AC ; Monsigny, M  (1979)

Lymphocytes membrane lectins - direct visualization by the use of glycosylated cytochemical markers

Biologie Cellulaire 36 (3) A5-A5

Kieda, C ; Roche, AC ; Delmotte, F ; Monsigny, M  (1979)

Lymphocyte membrane lectins - direct visualization by the use of fluoresceinyl-glycosylated cytochemical markers

Febs Letters 99 (2) 329-332

Schrevel, J ; Kieda, C ; Caigneaux, E ; Gros, D ; Delmotte, F ; Monsigny, M  (1979)

Visualization of cell surface carbohydrates by a general two-step lectin technique : lectins and glycosylated cytochemical markers.

Biologie Cellulaire 36 (3) 259-266

Rougier, M ; Kieda, C ; Monsigny, M  (1979)

Use of lectin to detect the sugar components of maize root cap slime

Journal of Histochemistry & Cytochemistry 27 (4) 878-881

Debray, H ; Decout, D ; Strecker, G ; Montreuil, J ; Monsigny, M  (1979)

Studies on the specificities of some lectins

Biologie Cellulaire 36 (3) A3-A3

Helene, C ; Montenay-Garestier, T ; Monsigny, M  (1979)

Some applications of low-temperature luminescence to the study of protein interactions with nucleic-acids and sugars

Journal of Luminescence 18-9 577-581

Monsigny, M ; Sene, C ; Obrenovitch, A  (1979)

Quantitative fluorimetric determination of cell-surface glycoconjugates with fluorescein-substituted lectins

European Journal of Biochemistry 96 (2) 295-300

Monsigny, M ; Sene, C ; Obrenovitch, A  (1979)

Quantitative fluorimetric determination of cell-surface glycoconjugates with fluorescein substituted lectins

Biologie Cellulaire 36 (3) A7-A7

Roche, AC. ; Monsigny, M  (1979)

Limulin (Limulus polyphemus lectin). Isolation, physicochemical properties, sugar specificity and mitogenic activity.

Progress in Clinical and Biological Research 29 603-616

Monsigny, M ; Schrevel, J  (1979)

International colloquim on membrane glycoconjugates - foreword

Biologie Cellulaire 36 (3) 209-212

Maget-Dana, R ; Roche, A C ; Monsigny, M  (1979)

Ganglioside-limulin interactions.

Progress in Clinical and Biological Research 295 67-78

Rougier, M ; Chaboud, A ; Kieda, C ; Monsigny, M  (1979)

Distribution and ultrastructural-localization of fucose-containing glycoconjugates at the surface of slime secretory root cap cells

Biologie Cellulaire 36 (3) A9-A9


1978   Références trouvées : 8

Roche, AC ; Magetdana, R ; Obrenovitch, A ; Hildenbrand, K ; Nicolau, C ; Monsigny, M  (1978)

Interaction between vesicles containing gangliosides and limulin (limulus-polyphemus lectin) - fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene

Febs Letters 93 (1) 91-96

Roche, AC ; Maurizot, JC ; Monsigny, M  (1978)

Circular-dichroism of limulin - limulus-polyphemus lectin

Febs Letters 91 (2) 233-236

Cohen, J ; Magetdana, R ; Roche, AC ; Monsigny, M  (1978)

Calf rotavirus - detection of outer capsid glycoproteins by lectins

Febs Letters 87 (1) 26-30

Redziniak, G ; Leclecr, M ; Panijel, J ; Monsigny, M  (1978)

Separation of 2 different populations of axial organ cells of asterias-rubens by use of lectins

Biochimie 60 (5) 525-527

Monsigny, M ; Jeune-Chung, KH ; Perrodon, Y  (1978)

Separation and biological properties of phaseolus-vulgaris isolectins

Biochimie 60 (11-1) 1315-1322

Sene, C ; Genest, D ; Obrenovitch, A ; Wahl, P ; Monsigny, M  (1978)

Pulse fluorimetry of 1,6-diphenyl-1,3,5-hexatriene incorporated in membranes of mouse leukemic-l-1210 cells

Febs Letters 88 (2) 181-186

Grivet, JP ; Delmotte, F ; Monsigny, M  (1978)

Protein-sugar interactions - nuclear magnetic-resonance investigation of binding of O-methyl-di-N-acetyl-ß-chitobioside to wheat-germ agglutinin (lectin)

Febs Letters 88 (2) 176-180

Obrenovitch, A ; Sene, C ; Negre, MT ; Monsigny, M  (1978)

Fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene embedded in membranes of mouse leukemic l-1210 cells during cell-cycle

Febs Letters 88 (2) 187-191


1977   Références trouvées : 7

Roche, AC ; Perrodon, Y ; Halpern, B ; Monsigny, M  (1977)

Limulin (limulus-polyphemus lectin) - mitogenic effect on human peripheral lymphocytes

European Journal of Immunology 7 (5) 263-267

Redziniak, G ; Leclerc, M ; Monsigny, M  (1977)

Visualization of membrane glycoconjugates of axial organ cells of asterias-rubens (asteridae, echinoderma) .1.

Biologie Cellulaire 30 (1) A18-A18

Gros, D ; Obrenovitch, A ; Challice, CE ; Monsigny, M ; Schrevel, J  (1977)

Ultrastructural visualization of cellular carbohydrate components by means of lectins on ultrathin glycol methacrylate sections

Journal of Histochemistry & Cytochemistry 25 (2) 104-114

Monsigny, MLP ; Delay, D ; Vaculik, M  (1977)

Synthesis of new type of glycoconjugate - thio-ß-D-glucopyranoside of l-cysteine

Carbohydrate Research 59 (2) 589-593

Obrenovitch, A ; Sene, C ; Negre, MT ; Monsigny, M  (1977)

Study on dynamics of plasma-membranes (l 1210) of mouse leukemia-cells as function of cellular cycle

Biologie Cellulaire 30 (1) A14-A14

Kieda, C ; Delmotte, F ; Monsigny, M  (1977)

Preparation and properties of glycosylated cytochemical markers

Febs Letters 76 (2) 257-261

Maget-Dana, R ; Roche, AC ; Monsigny, M  (1977)

Interaction between vesicles containing gangliosides and lectins - Limuklin and wheat-germ agglutinin

Febs Letters 79 (2) 305-309


1976   Références trouvées : 6

Gros, D ; Challice, CE ; Monsigny, M ; Schrevel, J  (1976)

Use of concanavallin-a on GMA sections for ultrastructural detection of extracellular and intracellular glucides

Journal de Microscopie et de Biologie Cellulaire 26 (2-3) A15-A15

Schrevel, J ; Kieda, C ; Gros, D ; Obrenovitch, A ; Caigneaux, E ; Delmotte, F ; Monsigny, M  (1976)

New tracers for membrane glycoconjugates - glycosylated peroxidase and ferritin

Journal de Microscopie et de Biologie Cellulaire 27 (1) A23-A23

Privat, JP ; Wahl, P ; Monsigny, M ; Auchet, JC  (1976)

Nanosecond-pulse fluorimetry of wheat-germ agglutinin (lectin)

European Journal of Biochemistry 68 (2) 573-580

Bouchard, P ; Moroux, Y ; Tixier, R ; Privat, JP ; Monsigny, M  (1976)

Improved method for purification of wheat-germ agglutinin (lectin) by affinity chromatography

Biochimie 58 (10) 1247-1253

Delmotte, F ; Kieda, C ; Monsigny, M  (1976)

Evidence and characterization of an endo-ß-N-acetyl glucosaminidase in rabbit blood-serum

Comptes Rendus Hebdomadaires Des Seances de L Academie Des Sciences Serie D 283 (9) 1121-1124

Privat, JP ; Lotan, R ; Bouchard, P ; Sharon, N ; Monsigny, M  (1976)

Chemical modification of tryptophan residues of wheat-germ agglutinin - effect on fluorescence and saccharide-binding properties

European Journal of Biochemistry 68 (2) 563-572


1975   Références trouvées : 6

Roche, AC ; Schauer, R ; Monsigny, M  (1975)

Protein-sugar interactions - purification by affinity chromatography of limulin, an n-acyl-neuraminidyl-binding protein

Febs Letters 57 (3) 245-249

Delmotte, F ; Kieda, C ; Monsigny, M  (1975)

Protein-sugar interaction - purification by affinity chromatography of solanum-tuberosum agglutinin (sta-lectin)

Febs Letters 53 (3) 324-330

Privat, JP ; Monsigny, M  (1975)

Luminescence studies of saccharide binding to wheat-germ agglutinin (lectin)

European Journal of Biochemistry 60 (2) 555-567

Cornet, M ; Alliot, F ; Pessac, B ; Monsigny, M  (1975)

Intercellular-adhesion of neuroretina chick-embryo cells - enhancement by bovine serum-albumin and derivates

Febs Letters 60 (2) 290-293

Delmotte, FM ; Privat, JPDJ ; Monsigny, MLP  (1975)

Glycan-protein interactions - synthesis of 4-methylumbelliferyl-2-acetamido-2-deoxy-ß-D-glucopyranoside, 4-methylumbelliferyl-di-N-acetyl-ß-chitobioside and 4-methyl-tri-N-acetyl-ß-chitotrioside - interaction of these osides with lysozyme

Carbohydrate Research 40 (2) 353-364

Cornet, M ; Alliot, F ; Monsigny, M ; Pessac, B  (1975)

Enhancement of intercellular-adhesion of chick-embryo neuro retinal cells by peptides from bovine serum-albumin

Comptes Rendus Hebdomadaires Des Seances de L Academie Des Sciences Serie D 281 (21) 1621-1623


1974   Références trouvées : 8

Rafestin, ME ; Delay, D ; Monsigny, M  (1974)

Synthesis of a Ll-fucopyranosides

Canadian Journal of Chemistry-Revue Canadienne de Chimie 52 (2) 210-212

Zachowski, A ; Prigent, B ; Monsigny, M ; Paraf, A  (1974)

Susceptible and non susceptible phenotypes of mopc 173 plasmocytoma to killing effect of concanavalin-a - study on concanavalin-a binding-sites

Biochimie 56 (11-1) 1621-1625

Rafestin, ME ; Obrenovitch, A ; Oblin, A ; Monsigny, M  (1974)

Purification of N-acetyl deuterium-glucosamine-binding proteins by affinity chromatography

Febs Letters 40 (1) 62-66

Privat, JP ; Delmotte, F ; Monsigny, M  (1974)

Protein-sugar interactions - association of wheat-germ agglutinin (lectin) and O-(4-methyl-umbelliferyl)-glycosides

Febs Letters 46 (1) 229-232

Privat, JP ; Delmotte, F ; Monsigny, M  (1974)

Protein-sugar interactions - association of ß-(1-]4) linked N-acetyl-D-glucosamine oligomer derivatives with wheat-germ agglutinin (lectin)

Febs Letters 46 (1) 224-228

Takerkart, G ; Segard, E ; Monsigny, M  (1974)

Preparation and properties of organophilic trypsin macroinhibitors - diamidino-a, ?-diphenylcarbamyl-poly(ethylene glycol)

Febs Letters 42 (2) 214-217

Takerkart, G ; Segard, E ; Monsigny, M  (1974)

Partition of trypsin in 2-phase systems containing a diamidino-a, ?-diphenylcarbamyl poly (ethylene-glycol) as competitive inhibitor of trypsin

Febs Letters 42 (2) 218-220

Privat, JP ; Delmotte, F ; Mialonier, G ; Bouchard, P ; Monsigny, M  (1974)

Fluorescence studies of saccharide binding to wheat-germ agglutinin (lectin)

European Journal of Biochemistry 47 (1) 5-14


1973   Références trouvées : 1

Mialonier, G ; Privat, JP ; Monsigny, M ; Kahlem, G ; Durand, R  (1973)

Isolation, physicochemical properties and in vivo localization of a phytohemagglutin (lectin) from phaseolus-vulgaris l (var red)

Physiologie Vegetale 11 (3) 519-537


1972   Références trouvées : 1

Monsigny M ; Delay, D  (1972)

Synthesis and properties of glyco peptides

Bulletin de la Societe Chimique de France 18b14


1970   Références trouvées : 1

Grimmonprez, L ; Bouquelet, S ; Bayard, B ; Spik, G ; Monsigny, M ; Montreuil, J  (1970)

Structure of hexasaccharide isolated from human milk - di-N-acetylneuraminyl tetrasaccharide

European Journal of Biochemistry 13 (3) 484-&


1969   Références trouvées : 8

Spik, G ; Monsigny, M ; Montreuil, J  (1969)

Study of glycoproteins. XXV. The C-terminal amino acids of human transferrin and lactotrans-ferrin.

Bulletin de la Societe de Chimie Biologique 50 (11) 2186-87
Human transferrin and lactotransferrin were heated with apotransferrin and lactoapotransferrin respectively using nor-leucine as internal standard, then heated with anhydrous hydrazine. Amino acids were separated on Amberlite IRC 50 and identified using a Technicon AutoAnalyzer. Number of C terminal amino acid residues/mole of protein of 76 000 mol. wt. in transferrin and lactotransferrin respectively were : glycine 0.80 and 1 ; proline 0.72 and 0 ; serine 0.32 and 0.60. SPT.

Human transferrin and lactotransferrin were heated with apotransferrin and lactoapotransferrin respectively using nor-leucine as internal standard, then heated with anhydrous hydrazine. Amino acids were separated on Amberlite IRC 50 and identified using a Technicon AutoAnalyzer. Number of C terminal amino acid residues/mole of protein of 76 000 mol. wt. in transferrin and lactotransferrin respectively were : glycine 0.80 and 1 ; proline 0.72 and 0 ; serine 0.32 and 0.60. SPT.

Monsigny, M ; Charet, P ; Spik, G ; Montreuil, J  (1969)

Structure of human transferrin-peptides and glycopeptides obtained by trypsin hydrolysis

Archives Internationales de Physiologie et de Biochimie 77 (3) 570-&

Jacubza, E ; Charet, P ; Monsigny, M ; Montreuil, J  (1969)

Solution to problems of ovomucoid heterogeneity by identification of its constituents

Hoppe-Seylers Zeitschrift Fur Physiologische Chemie 350 (1) 6-&

Montreuil, J ; Monsigny, M ; Spik, G ; Bayard, B  (1969)

Solution of some problems in glycan structure

Hoppe-Seylers Zeitschrift Fur Physiologische Chemie 350 (6) 667-&

Montreuil, J ; Monsigny, M ; Spik, G ; Duquesne, N ; Cheron, A ; Descamps, J ; Fournet, B  (1969)

Methods for determining glucide-peptide bonds in glycoproteins . An hypothesis of peptide chain encoding from points of glycan conjugation

Hoppe-Seylers Zeitschrift Fur Physiologische Chemie 350 (6) 664-&

Descamps, J ; Monsigny, M ; Montreuil, J  (1969)

Glycoprotein bonds in ?-A-globulin in human milk

Hoppe-Seylers Zeitschrift Fur Physiologische Chemie 350 (1) 4-&

Charet, P ; Monsigny, M ; Spik, G ; Montreuil, J  (1969)

Glycoproteins and peptide sequences of 2 glycopeptides isolated from trypsin hydrolysates of human transferrin

Comptes Rendus Hebdomadaires Des Seances de L Academie Des Sciences Serie D 269 (11) 1019-&

Monsigny, M  (1969)

Glycoproteins .37. Description of an automatic analyser of osamines adapted to column chromatography with cation exchange resins

Bulletin de la Societe de Chimie Biologique 51 (9) 1263-&


1968   Références trouvées : 6

Montreuil, J ; Takerkart, G ; Monsigny, M ; Bayard, B ; Dupony, P ; Fournet, B ; Grimmonprez, L  (1968)

Structural exploration of glucide groupings of glycoproteins (partial degradation by hydrolysis and acetolysis - permethylation)

Archives de Biochimie et Cosmetologie 11 (111) 26-&

Descamps, J ; Monsigny, M ; Montreuil, J  (1968)

Glycoprotein research . Discovery of O-seryl and O-threonyl-N-acetylgalactosaminidic bonds in ?-A globulins of human breast milk

Comptes Rendus Hebdomadaires Des Seances de L Academie Des Sciences Serie D 266 (17) 1775-&

Monsigny, M  (1968)

Glycoproteins .26. Simultaneous automatic amino acid glucosamine and galactosamine assay in glycoprotein hydrolysates

Bulletin de la Societe de Chimie Biologique 50 (11) 2188-&

Spik, G ; Monsigny, M ; Montreuil, J  (1968)

Glycoproteins .25. C-terminal amino acids of human transferrin and lactotransferrin

Bulletin de la Societe de Chimie Biologique 50 (11) 2186-&

Monsigny, M ; Adam-Chosson, A ; Montreuil, J  (1968)

Glycoproteins .22. Determining nature of point of glycanne-protein binding in chicken ovomucoid preparations

Bulletin de la Societe de Chimie Biologique 50 (4) 857-&

Monsigny, M ; Spik, G ; Montreuil, J  (1968)

Automatic column chromatography of glucosamine and galactosamine in glycoprotein hydrolysates

Archives de Biochimie et Cosmetologie 11 (111) 25-&


1967   Références trouvées : 3

Grimmonprez, L ; Takerkart, G ; Monsigny, M ; Montreuil, J  (1967)

Structure of an isolated human-milk hexose - di-lactaminyl-lacto-N-tetraose

Comptes Rendus Hebdomadaires Des Seances de L Academie Des Sciences Serie D 265 (25) 2124-&

Grimmonprez, L ; Takerkart, G ; Monsigny, . ; Montreuil, J  (1967)

Structure of a hexasaccharide isolated from human milk : dilactaminyl-lacto-N-tetraose.

Comptes Rendus Hebdomadaires Des Seances de L’academie Des Sciences Serie D 265d 25 2124-26
The hexasaccharide was subjected to partial acid hydrolysis (0.1N H2SO4, 80 deg C), further hydrolysis (0.5N, 100 deg C), oxidation by periodic acid and methylation. In each case the products were passed through an ion-exchange column and identified by paper chromatography. The hexasaccharide consisted of D-galactopyranose, N-acetyl-o-pyranoglucosamine, D-glucopyranose and N-acetyl-neuraminic acid in the proportions 2 : 1 : 1 : 2. The structure was that of lacto-N-tetraose (see DSA 25 : 447) with the addition of 2 neuraminic acid residues on position 3 of the terminal galactose and position 6 of the N-acetyl-glucosamine. JK.

The hexasaccharide was subjected to partial acid hydrolysis (0.1N H2SO4, 80 deg C), further hydrolysis (0.5N, 100 deg C), oxidation by periodic acid and methylation. In each case the products were passed through an ion-exchange column and identified by paper chromatography. The hexasaccharide consisted of D-galactopyranose, N-acetyl-o-pyranoglucosamine, D-glucopyranose and N-acetyl-neuraminic acid in the proportions 2 : 1 : 1 : 2. The structure was that of lacto-N-tetraose (see DSA 25 : 447) with the addition of 2 neuraminic acid residues on position 3 of the terminal galactose and position 6 of the N-acetyl-glucosamine. JK.

Montreuil, J ; Monsigny, M ; Buchet, MT  (1967)

Etudes sur les glycoprotéides . action des alcalis sur les O-seryl et O-threonyl-ß-N-acetyl-D-glucosaminides . Mécanisme de la ß-élimination

Comptes Rendus Hebdomadaires Des Seances de L Academie Des Sciences Serie D 264 (16) 2068-&


1966   Références trouvées : 4

Monsigny, M ; Montreuil, J  (1966)

Studies on glycoproteins. Demonstration of a binding of aspartic acid and glucosamine and an O-threonyl-glycoside binding in human lactotransferrin

Comptes Rendus Hebdomadaires Des Seances de L Academie Des Sciences. D : Sciences Naturelles 263 (12) 893-5

Monsigny, M ; Montreuil, J  (1966)

Etude sur les glycoprotéides . Nature du point d’attache glycanne-protéine dans lovomucoide

Comptes Rendus Hebdomadaires Des Seances de L Academie Des Sciences Serie D 262 (16) 1780-&

Duquesne, N ; Monsigny, M ; Montreuil, J  (1966)

Etudes sur les glycoprotéides . Nature du point d’attache glycanne-protéine dans les globulines ? G du serum humain

Comptes Rendus Hebdomadaires Des Seances de L Academie Des Sciences Serie D 262 (24) 2536-&

Spik, G ; Monsigny, M ; Montreuil, J  (1966)

Etudes sur les glycoprotéides . Mise en évidence dune liaison de lacide aspartique et de la glucosamine et dune liaison O-threonyl-glycosidique dans la lactotransferrine humaine

Comptes Rendus Hebdomadaires Des Seances de L Academie Des Sciences Serie D 263 (12) 893-&


1965   Références trouvées : 8

Montreuil, J ; Spik, G ; Dumaisnil, J ; Monsigny, M  (1965)

Procédés de determination de la composition en oses des osides libres et combines

Bulletin de la Societe Chimique de France (1) 239-&

Duquesne, N ; Monsigny, M ; Montreuil, J  (1965)

Etudes sur les glycoprotéides . Nature du point d’attache glycanne-protéine dans les globulines ? G du serum de boeuf

Pathologie Biologie 13 (23-2) 1246-&

Duquesne, N ; Monsigny, M ; Montrueil, J  (1965)

Etudes sur les glycoproteides. Nature du point d’attache glycanne-protéine dans les globulines ? G du serum de boeuf

Comptes Rendus Hebdomadaires Des Seances de L Academie Des Sciences 261 (5) 1430-&

Spik, G ; Monsigny, M ; Montreuil, J  (1965)

Etudes sur les glycoprotéides . Mise en évidence dune liaison de lacide aspartique et de la glucosamine dans la transferrine humaine

Pathologie Biologie 13 (23-2) 1244-&

Spik, G ; Monsigny, M ; Montreuil, J  (1965)

Etude sur les glycoprotéides . Mise en évidence dune liaison de lacide aspartique avec le groupement mucopolyosidique dans la transferrine humaine

Pathologie Biologie 13 (15-1) 844-&

Spik, G ; Monsigny, M ; Montreuil ; J  (1965)

Etudes sur les glycoprotéides .Mise en évidence dune liaison de lacide aspartique et de la glucosamine dans la transferrine humaine

Comptes Rendus Hebdomadaires Des Seances de L Academie Des Sciences 261 (4) 1137-&

Spik, G ; Monsigny, M ; Montreuil, J  (1965)

Etude comparée de la structure de la transferrine et de la lactotransferrine humaines

Bulletin de la Societe de Chimie Biologique 47 (12) 2329-&

Montreuil, J ; Spik, G ; Monsigny, M ; Descamps, J ; Biserte, G ; Dautrevaux, M  (1965)

Etude comparée de la composition en oses et en amino-acides de la transferrine et de la lactotransferrine humaines

Experientia 21 (5) 254-&


Mots-clés

Professeur émérite , Microenvironnement cellulaire et cibles pharmacologiques