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Blood, Vol. 94 No. 3 (August 1), 1999:
pp. 1012-1020
Surface Molecule Loss and Bleb Formation by Human Germinal Center B
Cells Undergoing Apoptosis: Role of Apoptotic Blebs in Monocyte
Chemotaxis
Carmen Segundo,
Francisco Medina,
Carmen Rodríguez,
Rosalía Martínez-Palencia,
Francisco Leyva-Cobián, and
José A. Brieva
From Servicio de Inmunología, Hospital Universitario Puerta
del Mar, Cádiz; and Servicio de Inmunología, Hospital
Universitario Marqués de Valdecilla, Santander, Spain.
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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