Uptake of plasmid/glycosylated polymer complexes and gene transfer efficiency in differentiated airway epithelial cells
Identifieur interne : 000361 ( France/Analysis ); précédent : 000360; suivant : 000362Uptake of plasmid/glycosylated polymer complexes and gene transfer efficiency in differentiated airway epithelial cells
Auteurs : Isabelle Fajac [France] ; Guiti Thévenot [France] ; Laurent Bédouet [France] ; Claire Danel [France] ; Marc Riquet [France] ; Marc Merten [France] ; Catherine Figarella [France] ; Josette Dall' Ava-Santucci [France] ; Michel Monsigny [France] ; Pascale Briand [France]Source :
- The Journal of Gene Medicine [ 1099-498X ] ; 2003-01.
English descriptors
- Teeft :
- Acidic compartments, Airway, Airway epithelial cells, Airway epithelium, Airway gland serous cells, Airway surface, Airway surface epithelial cells, Attened cells, Bicinchoninic acid, Biol, Biol chem, Bronchial tissue, Brosis, Cationic, Cationic lipids, Cationic polymer, Cell culture, Cell culture model, Cell fluorescence intensity, Cellular uptake, Cfkm4 cells, Chloroquine, Complexed, Copyright, Culture medium, Cystic, Cytokeratin, Endocytosis, Endocytotic properties, Endosomolytic agents, Epithelial, Epithelium, Explant, Fajac, Gene, Gene therapy, Gene transfer, Glycosylated, Glycosylated complexes, Glycosylated polylysine, Glycosylated polylysines, Glycosylated polymers, Human airway surface epithelium, Hypertonic, Hypertonic medium, Intracellular, John wiley sons, Lactose, Lactosylated, Lactosylated polylysine, Lectin, Leitz microscope, Luciferase, Mannose, Mannosylated, Membrane lectins, Midoux, Molecular probes, Monensin, Neoglycoproteins, Outgrowth, Plasmid, Polyethylenimine, Polylysine, Polylysines, Polymer, Receptor, Results show, Roche, Room temperature, Serous, Similar amounts, Sodium pyruvate, Synthetic vectors, Transfected, Transfected cells, Transfection, Ultroser, Undifferentiated cells, Unsubstituted, Uoresceinated, Uorescence, Uorescence intensity, Uorescent, Uorescent cells, Uptake.
Abstract
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. Methods: In both cell types, lactosylated PEI was more efficient for gene transfer than unsubstituted PEI and lactosylated polylysine which requires the presence of endosomolytic agents. However, for all the vectors tested, gene transfer efficiency was lower in differentiated cells as compared with poorly differentiated cells. The presence of membrane lectins, i.e. cell surface sugar‐specific receptors, was evaluated using fluorescein‐conjugated neoglycoproteins and microscopy or flow cytometry. In differentiated airway surface epithelial cells, membrane lectins were not expressed and plasmid DNA/fluorescein‐conjugated glycosylated polymer complexes were not incorporated. This accounted in part for the lack of gene transfer efficiency in these cells. In contrast, in differentiated airway gland serous cells, expression of lectins and their endocytotic properties appeared to be similar to that observed in undifferentiated cells, and plasmid DNA/fluorescein‐conjugated glycosylated polymer complexes were incorporated in similar amounts by cells in both differentiated states. Results: Glycosylated PEI appears to be a promising gene delivery system since it is more efficient than the sugar‐free polymer and does not require endosomolytic agents. However, in differentiated airway gland serous cells, a low gene transfer efficiency was observed that could not be attributed to low expression of membrane lectins or low uptake of glycosylated complexes. An impaired intracellular trafficking of glycosylated complexes in differentiated airway gland serous cells is suggested. Copyright © 2002 John Wiley & Sons, Ltd.
Url:
DOI: 10.1002/jgm.318
Affiliations:
- France
- Centre-Val de Loire, Grand Est, Lorraine (région), Provence-Alpes-Côte d'Azur, Région Centre, Île-de-France
- Marseille, Nancy, Orléans, Paris
- Université d'Orléans
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ISTEX:63B860374DDF2983581D140BD87877694015D1CALe document en format XML
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first="Isabelle" last="Fajac">Isabelle Fajac</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>Laboratoire de Physiologie Respiratoire, CHU Cochin, AP‐HP‐Université Paris V, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation><affiliation wicri:level="1"><country wicri:rule="url">France</country></affiliation><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>Correspondence address: Laboratoire de Physiologie Respiratoire, Faculté Cochin, 24 rue du fg St Jacques, 75014 Paris</wicri:regionArea><placeName><region type="region" nuts="2">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Thevenot, Guiti" sort="Thevenot, Guiti" uniqKey="Thevenot G" first="Guiti" last="Thévenot">Guiti Thévenot</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>Laboratoire de Physiologie Respiratoire, CHU Cochin, AP‐HP‐Université Paris V, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Bedouet, Laurent" sort="Bedouet, Laurent" uniqKey="Bedouet L" first="Laurent" last="Bédouet">Laurent Bédouet</name><affiliation wicri:level="4"><country xml:lang="fr">France</country><wicri:regionArea>Glycobiologie, Centre de Biophysique Moléculaire, CNRS et Université d'Orléans, Orléans</wicri:regionArea><placeName><region type="region">Centre-Val de Loire</region><region type="old region">Région Centre</region><settlement type="city">Orléans</settlement></placeName><orgName type="university">Université d'Orléans</orgName></affiliation></author><author><name sortKey="Danel, Claire" sort="Danel, Claire" uniqKey="Danel C" first="Claire" last="Danel">Claire Danel</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>Service d'Anatomie‐Pathologie, Hôpital Européen Georges Pompidou, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Riquet, Marc" sort="Riquet, Marc" uniqKey="Riquet M" first="Marc" last="Riquet">Marc Riquet</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>Service de Chirurgie Thoracique, Hôpital Européen Georges Pompidou, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Merten, Marc" sort="Merten, Marc" uniqKey="Merten M" first="Marc" last="Merten">Marc Merten</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>INSERM 00‐14, Faculté de Médecine, Nancy</wicri:regionArea><placeName><region type="region">Grand Est</region><region type="old region">Lorraine (région)</region><settlement type="city">Nancy</settlement></placeName></affiliation></author><author><name sortKey="Figarella, Catherine" sort="Figarella, Catherine" uniqKey="Figarella C" first="Catherine" last="Figarella">Catherine Figarella</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>Groupe de Recherche sur les Glandes Exocrines, Faculté de Médecine, Marseille</wicri:regionArea><placeName><region type="region">Provence-Alpes-Côte d'Azur</region><region type="old region">Provence-Alpes-Côte d'Azur</region><settlement type="city">Marseille</settlement></placeName></affiliation></author><author><name sortKey="Ava Antucci, Josette Dall" sort="Ava Antucci, Josette Dall" uniqKey="Ava Antucci J" first="Josette Dall'" last="Ava-Santucci">Josette Dall' Ava-Santucci</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>Laboratoire de Physiologie Respiratoire, CHU Cochin, AP‐HP‐Université Paris V, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author><author><name sortKey="Monsigny, Michel" sort="Monsigny, Michel" uniqKey="Monsigny M" first="Michel" last="Monsigny">Michel Monsigny</name><affiliation wicri:level="4"><country xml:lang="fr">France</country><wicri:regionArea>Glycobiologie, Centre de Biophysique Moléculaire, CNRS et Université d'Orléans, Orléans</wicri:regionArea><placeName><region type="region">Centre-Val de Loire</region><region type="old region">Région Centre</region><settlement type="city">Orléans</settlement></placeName><orgName type="university">Université d'Orléans</orgName></affiliation></author><author><name sortKey="Briand, Pascale" sort="Briand, Pascale" uniqKey="Briand P" first="Pascale" last="Briand">Pascale Briand</name><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>Institut Cochin, INSERM U567, Université Paris V, Paris</wicri:regionArea><placeName><region type="region">Île-de-France</region><region type="old region">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">The Journal of Gene Medicine</title><title level="j" type="alt">JOURNAL OF GENE MEDICINE, THE</title><idno type="ISSN">1099-498X</idno><idno type="eISSN">1521-2254</idno><imprint><biblScope unit="vol">5</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="38">38</biblScope><biblScope unit="page" to="48">48</biblScope><biblScope unit="page-count">11</biblScope><publisher>John Wiley & Sons, Ltd.</publisher><pubPlace>Chichester, UK</pubPlace><date type="published" when="2003-01">2003-01</date></imprint><idno type="ISSN">1099-498X</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1099-498X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Acidic compartments</term><term>Airway</term><term>Airway epithelial cells</term><term>Airway epithelium</term><term>Airway gland serous cells</term><term>Airway surface</term><term>Airway surface epithelial cells</term><term>Attened cells</term><term>Bicinchoninic acid</term><term>Biol</term><term>Biol chem</term><term>Bronchial tissue</term><term>Brosis</term><term>Cationic</term><term>Cationic lipids</term><term>Cationic polymer</term><term>Cell culture</term><term>Cell culture model</term><term>Cell fluorescence intensity</term><term>Cellular uptake</term><term>Cfkm4 cells</term><term>Chloroquine</term><term>Complexed</term><term>Copyright</term><term>Culture medium</term><term>Cystic</term><term>Cytokeratin</term><term>Endocytosis</term><term>Endocytotic properties</term><term>Endosomolytic agents</term><term>Epithelial</term><term>Epithelium</term><term>Explant</term><term>Fajac</term><term>Gene</term><term>Gene therapy</term><term>Gene transfer</term><term>Glycosylated</term><term>Glycosylated complexes</term><term>Glycosylated polylysine</term><term>Glycosylated polylysines</term><term>Glycosylated polymers</term><term>Human airway surface epithelium</term><term>Hypertonic</term><term>Hypertonic medium</term><term>Intracellular</term><term>John wiley sons</term><term>Lactose</term><term>Lactosylated</term><term>Lactosylated polylysine</term><term>Lectin</term><term>Leitz microscope</term><term>Luciferase</term><term>Mannose</term><term>Mannosylated</term><term>Membrane lectins</term><term>Midoux</term><term>Molecular probes</term><term>Monensin</term><term>Neoglycoproteins</term><term>Outgrowth</term><term>Plasmid</term><term>Polyethylenimine</term><term>Polylysine</term><term>Polylysines</term><term>Polymer</term><term>Receptor</term><term>Results show</term><term>Roche</term><term>Room temperature</term><term>Serous</term><term>Similar amounts</term><term>Sodium pyruvate</term><term>Synthetic vectors</term><term>Transfected</term><term>Transfected cells</term><term>Transfection</term><term>Ultroser</term><term>Undifferentiated cells</term><term>Unsubstituted</term><term>Uoresceinated</term><term>Uorescence</term><term>Uorescence intensity</term><term>Uorescent</term><term>Uorescent cells</term><term>Uptake</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">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. Methods: In both cell types, lactosylated PEI was more efficient for gene transfer than unsubstituted PEI and lactosylated polylysine which requires the presence of endosomolytic agents. However, for all the vectors tested, gene transfer efficiency was lower in differentiated cells as compared with poorly differentiated cells. The presence of membrane lectins, i.e. cell surface sugar‐specific receptors, was evaluated using fluorescein‐conjugated neoglycoproteins and microscopy or flow cytometry. In differentiated airway surface epithelial cells, membrane lectins were not expressed and plasmid DNA/fluorescein‐conjugated glycosylated polymer complexes were not incorporated. This accounted in part for the lack of gene transfer efficiency in these cells. In contrast, in differentiated airway gland serous cells, expression of lectins and their endocytotic properties appeared to be similar to that observed in undifferentiated cells, and plasmid DNA/fluorescein‐conjugated glycosylated polymer complexes were incorporated in similar amounts by cells in both differentiated states. Results: Glycosylated PEI appears to be a promising gene delivery system since it is more efficient than the sugar‐free polymer and does not require endosomolytic agents. However, in differentiated airway gland serous cells, a low gene transfer efficiency was observed that could not be attributed to low expression of membrane lectins or low uptake of glycosylated complexes. An impaired intracellular trafficking of glycosylated complexes in differentiated airway gland serous cells is suggested. Copyright © 2002 John Wiley & Sons, Ltd.</div></front></TEI><affiliations><list><country><li>France</li></country><region><li>Centre-Val de Loire</li><li>Grand Est</li><li>Lorraine (région)</li><li>Provence-Alpes-Côte d'Azur</li><li>Région Centre</li><li>Île-de-France</li></region><settlement><li>Marseille</li><li>Nancy</li><li>Orléans</li><li>Paris</li></settlement><orgName><li>Université d'Orléans</li></orgName></list><tree><country name="France"><region name="Île-de-France"><name sortKey="Fajac, Isabelle" sort="Fajac, Isabelle" uniqKey="Fajac I" first="Isabelle" last="Fajac">Isabelle Fajac</name></region><name sortKey="Ava Antucci, Josette Dall" sort="Ava Antucci, Josette Dall" uniqKey="Ava Antucci J" first="Josette Dall'" last="Ava-Santucci">Josette Dall' Ava-Santucci</name><name sortKey="Bedouet, Laurent" sort="Bedouet, Laurent" uniqKey="Bedouet L" first="Laurent" last="Bédouet">Laurent Bédouet</name><name sortKey="Briand, Pascale" sort="Briand, Pascale" uniqKey="Briand P" first="Pascale" last="Briand">Pascale Briand</name><name sortKey="Danel, Claire" sort="Danel, Claire" uniqKey="Danel C" first="Claire" last="Danel">Claire Danel</name><name sortKey="Fajac, Isabelle" sort="Fajac, Isabelle" uniqKey="Fajac I" first="Isabelle" last="Fajac">Isabelle Fajac</name><name sortKey="Fajac, Isabelle" sort="Fajac, Isabelle" uniqKey="Fajac I" first="Isabelle" last="Fajac">Isabelle Fajac</name><name sortKey="Figarella, Catherine" sort="Figarella, Catherine" uniqKey="Figarella C" first="Catherine" last="Figarella">Catherine Figarella</name><name sortKey="Merten, Marc" sort="Merten, Marc" uniqKey="Merten M" first="Marc" last="Merten">Marc Merten</name><name sortKey="Monsigny, Michel" sort="Monsigny, Michel" uniqKey="Monsigny M" first="Michel" last="Monsigny">Michel Monsigny</name><name sortKey="Riquet, Marc" sort="Riquet, Marc" uniqKey="Riquet M" first="Marc" last="Riquet">Marc Riquet</name><name sortKey="Thevenot, Guiti" sort="Thevenot, Guiti" uniqKey="Thevenot G" first="Guiti" last="Thévenot">Guiti Thévenot</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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