Cloning, expression and activity of B cell Activating Factor
Abstract
The preimmune antibody repertoire plays an important role in immune protection
through out the life of the organism. In rabbits, B cells develop in the bone marrow and
migrate to the gut associated lymphoid tissues (GALT), where in the presence of
commensal bacteria, they are stimulated to proliferate, form organized follicles and
develop -the primary antibody repertoire by Ig gene diversification.
While the molecular events leading to Ig gene diversification in the bone marrow are
well understood, little is known about the mechanisms regulating B cell proliferation and
homeostasis in GALT. In a previous study to test whether B cell activating factor (BAFF)
may play a role in stimulating GALT B cells, endogenous BAFF was neutralized in vivo
using a soluble decoy receptor (TACI-Ig). This treatment greatly reduced the number and
size of proliferating B cell follicles, demonstrating that BAFF is required for B cell
development in GALT. However, it was not clear from this study, whether BAFF
provided a proliferation signal or survival/maintenance signal. Therefore to address this
question, I cloned, expressed and purified recombinant rabbit BAFF from E. coli, and
tested its affect on B cells in vitro with a [3H]-thymidine incorporation assay. Purified
rabbit BAFF induced splenocytes to undergo proliferation, but not B cells isolated from
GALT. However, addition of BAFF to cultures of GALT B cells transiently prolonged
their survival, suggesting that BAFF may provide a survival/maintenance signal for B
cells in GALT.
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