Surface molecule loss and bleb formation by human germinal center B cells undergoing apoptosis: role of apoptotic blebs in monocyte chemotaxis

C Segundo, F Medina, C Rodrıguez… - Blood, The Journal …, 1999 - ashpublications.org
C Segundo, F Medina, C Rodrıguez, R Martınez-Palencia, F Leyva-Cobián, JA Brieva
Blood, The Journal of the American Society of Hematology, 1999ashpublications.org
Human tonsil germinal center (GC) B cells rapidly undergo apoptosis in culture. Annexin-V
binding shows an early event in this process. In the present study, this method has been
used to label apoptotic GC B cells and to analyze additional surface molecules. The
expression of all of the molecules studied was reduced in apoptotic (annexin-V+) GC B cells,
and the reduction was more marked for CD11a, CD21, CD22, CD49d, and CD54, molecules
that participate in survival interaction for GC B cells. The analysis of CD54, one of the …
Human tonsil germinal center (GC) B cells rapidly undergo apoptosis in culture. Annexin-V binding shows an early event in this process. In the present study, this method has been used to label apoptotic GC B cells and to analyze additional surface molecules. The expression of all of the molecules studied was reduced in apoptotic (annexin-V+) GC B cells, and the reduction was more marked for CD11a, CD21, CD22, CD49d, and CD54, molecules that participate in survival interaction for GC B cells. The analysis of CD54, one of the molecules that was more drastically reduced, showed that GC, but not mantle zone, B cells actively secrete CD54 to the culture supernatant (SN). The secreted CD54 was partly released from the GC B cells in a particulate form as demonstrated by centrifugation. Further experiments using filtration, fluorescence microscopy, electron microscopy, and flow cytometry analysis showed that GC B cells released to the culture SN a population of spherical membranous vesicles of about 0.18 μm in size, similar to the blebs described in other apoptosis systems. Bleb formation depended on active metabolism, Ca2+, and, in part, on microfilament integrity. GC B-cell–derived blebs were clearly associated with apoptosis, as antiapoptotic stimuli prevented their formation. In addition, GC B-cell–derived blebs contained the adhesion molecules previously studied. Consequently, bleb formation might contribute to the surface molecule loss occurring in apoptotic GC B cells. Finally, a chemotaxis assay showed that GC B-cell blebs were chemotactic for human monocytes, suggesting that this mechanism might operate in vivo.
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