Lectin-based three-color flow cytometric approach for studying cell surface glycosylation changes that occur during apoptosis
| Autor: | Jeanine Marquet, Yannick Pilatte, Anne Greffard, Chantal Batisse, Marie-Claude Jaurand, Jocelyne Fleury-Feith |
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| Rok vydání: | 2004 |
| Předmět: |
Glycosylation
Blotting Western Cell Biophysics Apoptosis Pathology and Forensic Medicine Flow cytometry chemistry.chemical_compound Endocrinology Agglutinin Annexin Lectins medicine Humans Propidium iodide biology medicine.diagnostic_test Cell Membrane Lectin U937 Cells Cell Biology Hematology Flow Cytometry Cell biology medicine.anatomical_structure chemistry biology.protein |
| Zdroj: | Cytometry. :81-88 |
| ISSN: | 1097-0320 0196-4763 |
| DOI: | 10.1002/cyto.a.20094 |
| Popis: | Background Changes in cell surface glycosylation that accompany apoptosis are thought to be involved in the recognition and removal of apoptotic cells by phagocytes, but in most instances these changes are ill defined. To improve our understanding of this phenomenon, we designed a trivariate flow cytometry procedure that allows direct comparison of cell surface glycosylation in apoptotic and viable cells Methods The annexin V/propidium iodide assay has been adapted for cell surface glycosylation analysis by combining the use of these two reagents with biotinylated lectins, and this has been used to investigate camptothecin-induced apoptosis in U-937 cells. Results Although numerous lectins are potent inducers of apoptosis, we found that it is possible to determine lectin concentrations that produce interpretable data without inducing significant cytotoxicity even when using apoptogenic lectins. That apoptosis is associated with a marked decrease in cell surface sialylation was confirmed by using the sialic acid–specific lectins Maackia amurensis agglutinin and Sambucus nigra agglutinin. These observations were corroborated by lectin blotting analysis with the same lectins. Conclusions Species- and cell-dependent altered glycosylation patterns are likely to be associated with different modes of apoptosis. The easy and versatile method described in this report should be useful for exploring this field. © 2004 Wiley-Liss, Inc. |
| Databáze: | OpenAIRE |
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