Membrane-bound CD154, but not CD40-specific antibody, mediates NF-kappaB-independent IL-6 production in B cells.

CD40, a member of the tumor necrosis factor receptor (TNF-R) superfamily, is expressed on the surface of B cells, where its engagement results in IL-6 secretion. In this study, we characterize the specific molecular requirements for CD40-mediated IL-6 production. Engagement of CD40 on either a B cell line or normal mouse splenic B cells with a membrane-bound form of CD154 (also known as CD40L or gp39) induced IL-6 secretion as well as up-regulation of IL-6 mRNA, but cross-linking CD40 with agonistic anti-CD40 mAb did not, although these mAb induce many other CD40 activation events, including the nuclear translocation of the transcription factor NF-kappaB. Using a mouse B cell line stably transfected with various human CD40 (hCD40) cytoplasmic truncation and point mutants, we show that the region from amino acids 202 to 225 in the cytoplasmic domain of CD40 is necessary for IL-6 secretion. However, the carboxy-terminal 32 amino acids are not, although these residues are required for CD40-mediated NF-kappaB activation. In addition, CD40 mutants previously shown to lack binding to TRAF2 and -3 are fully capable of inducing IL-6 production. Thus, CD40-mediated IL-6 induction is independent of NF-kappaB activation and the binding of TRAF2 and -3, but CD40 must be engaged by trimeric CD154 on cell membranes to activate production of IL-6.