Overexpression of BSAP/Pax-5 Inhibits Switching to IgA and Enhances Switching to IgE in the I.29μ B Cell Line

Abstract

AbstractB cell-specific activator protein (BSAP)/Pax-5 is a paired domain DNA-binding protein expressed in the developing nervous system, testis, and in all B lineage cells, except terminally differentiated plasma cells. BSAP regulates transcription of several genes expressed in B cells and also the activity of the 3′ IgH enhancer. As it has binding sites within or 5′ to the switch regions of nearly all Ig heavy chain C region genes and also is known to increase transcription of the germline ε RNA, BSAP has been hypothesized to be involved in regulation of Ab class switch recombination. To directly examine the effects of BSAP on isotype switching, we use a tetracycline-regulated expression system to overexpress BSAP in the surface IgM+ I.29μ B cell line, a mouse cell line that can be induced to undergo class switch recombination. We find that overexpression of BSAP inhibits switching to IgA in I.29μ cells stimulated with LPS + TGF-β1 + nicotinamide, but enhances switching to IgE in cells stimulated with LPS + IL-4 + nicotinamide. Parallel to its effects on switching, overexpression of BSAP inhibits germline α RNA expression and the transcriptional activity of the germline α promoter, while enhancing activity of the germline ε promoter. Proliferation of I.29μ cells is not affected in this system. The possible mechanisms and significance of the effect of BSAP on isotype switching are discussed.