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International Immunology, Vol. 5, No. 9, pp. 1085-1091,September 1993
© 1993 Japanese Society for Immunology

Distribution of cAMP in secondary follicles and its expression in B cell apoptosis and CD40-mediated survival

Kirstine A. Knox, Gerald D. Johnson and John Gordon

The Department of Immunology, The University of Birmingham Birmingham B15 2TJ, UK

Correspondence to: Correspondence to: K A. Knox

Apoptosis occurs at a high rate among B cells in germinal centres (GCs). Isolated GC B cells undergo apoptosis spontaneously when cultured in vitro: such self-destruction can be arrested by protein kinase C-activating phorbol esters and by ligating surface CD40. This study sought to explore whether the cAMP-dependent second messenger system played a role in the regulation of GC cell apoptosis and in ligand-promoted survival. First, the distribution of cAMP in GCs was analysed by immunofluorescence staining and confocal laser scanning microscopy in situ: cytoplasmlc cAMP was expressed by all cells but was most abundant in the constitutive cells of the light zones, where GC B cell apoptosis and rescue occur. Isolated GC B cells were found to express higher levels of cAMP than quiescent B lymphocytes as assessed both by a competitive binding assay and a newly developed flow cytometric method. Culturing GC B cells alone increased cAMP level, while those cells rescued with the phorbol ester phorbol myrlstate acetate (PMA) and anti-CD40 exhibited decreased cAMP levels. Resting B lymphocytes showed no change in cAMP level following culture alone, but a significant increase in cAMP levels when co-cultured with PMA and anti-CD40. These data suggest that given identical signals, resting B cells and GC B cells exhibit differential regulation of the cAMP-dependent second messenger system; moreover, this second messenger system appears to be involved in the regulation of apoptosis in GC B cells.

Keywords: centroblast, centrocyte, germinal centre, phorbol ester

Received 23 December 1992, accepted 24 May 1993.


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