Induction of Nuclear Factor-κB During Primary B Cell Differentiation

Abstract We have investigated activation of nuclear factor-κB (NF-κB) in the process of primary B cell differentiation in vitro. In this system, NF-κB is strongly induced when B cells develop from the pre-B cell to the immature B cell stage. Unlike the typical NF-κB activation in response to exogenous stimuli, induction proceeds with a slow time course. NF-κB induction is only observed in B cells that undergo differentiation, not in Rag2-deficient cells. Nuclear DNA binding complexes predominantly comprise p50/RelA heterodimers and, to a lesser extent, c-Rel-containing dimers. The increase in NF-κB binding activity is accompanied by a slow and steady decrease in IκBβ protein levels. Interestingly, absolute RelA protein levels remain unaffected, whereas RelB and c-Rel synthesis is induced. The reason for preferential nuclear translocation of RelA complexes appears to be selective inhibition by the IκBβ protein. IκBβ can efficiently inhibit p50/RelA complexes, but has a much reduced ability to interfere with p50/c-Rel DNA binding both in vitro and in vivo. Interestingly, p50/RelB complexes are not at all targeted by IκBβ, and coimmunoprecipitation experiments show no evidence for an association of IκBβ and RelB in vivo. Consistent with these observations, IκBβ cotransfection can inhibit p50/RelA-mediated trans-activation, but barely affects p50/RelB mediated trans-activation.