Although immunoglobulin (Ig)M hiIgD lo/−CD21 hi marginal zone B cells represent a significant proportion of naive peripheral splenic B lymphocytes, few of the genes that regulate their development have been identified. This subset of peripheral B cells fails to emerge in mice that lack nuclear factor (NF)-κBp50. Less drastic reductions in marginal zone B cell numbers are also seen in the spleens of recombination activating gene (Rag)-2 −/− mice reconstituted with NF-κBp65 −/− fetal liver cells and in c-Rel −/− mice. In contrast, steady-state levels of IgD hi splenic follicular B cells are not significantly reduced in the absence of NF-κBp50, NF-κBp65, or c-Rel. Reconstitution of B cells in Rag-2 −/− mice with a mixture of p50 −/−/p65 −/− fetal liver cells and Rag-2 −/− bone marrow cells revealed that the generation of marginal zone B cells requires the expression of NF-κB in developing B cells, as opposed to supporting cells.