The generation of VH81X heavy chain λ-light chain-expressing B cells (VH81X-λ+ B cells) was studied in VH81X heavy chain transgenic mice as well as in VH81X JH -/- and VH81X JH -/- Ck -/- mice, in which competition resulting from expression of heavy and light chains from the endogenous heavy and κ light chain loci was prevented. We show that although λ light chain gene rearrangements occur normally and give rise to light chains that associate with the transgenic heavy chain to form surface and soluble IgM molecules, further B cell development is almost totally blocked. The few VH81X-λ+ B cells that are generated progress into a mature compartment (expressing surface CD21, CD22, CD23, and low CD24 and having a relatively long life span) but they also have reduced levels of surface Ig receptor and express higher amounts of Fas Ag than VH81X-κ+ B cells. These VH81X-λ+ B cells reach the peripheral lymphoid organs and accumulate in the periarteriolar lymphoid sheath but are unable to generate primary B cell follicles. In other heavy chain transgenic mice (MD2, M167, and M54), λ+ B cells are generated. However, they seem to be preferentially selected in the peripheral repertoire of some transgenic heavy chain mice (M54) but not in others (MD2, M167). These studies show that a crucial selection step is necessary for B cell survival and maintenance in which B cells, similar to T cells, receive signals depending on their clonal receptors.