title: Lectin-based three-color flow cytometric approach for studying cell surface glycosylation changes that occur during apoptosis.
abstract: 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.
authors.affiliation: EMI 99-09 INSERM, Faculté de Médecine Paris XII, Créteil, France.
authors.forename: Chantal
authors.initials: C
authors.lastname: Batisse
authors.forename: Jeanine
authors.initials: J
authors.lastname: Marquet
authors.forename: Anne
authors.initials: A
authors.lastname: Greffard
authors.forename: Jocelyne
authors.initials: J
authors.lastname: Fleury-Feith
authors.forename: Marie-Claude
authors.initials: MC
authors.lastname: Jaurand
authors.forename: Yannick
authors.initials: Y
authors.lastname: Pilatte
mesh_terms: D017209:Apoptosis / Q000502:physiology*; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods*; D006031:Glycosylation; D006801:Humans; D037102:Lectins / Q000378:metabolism*; D020298:U937 Cells
chemical_list: D037102:Lectins