Antinuclear antibodies are present in the serum of individuals with systemic autoimmune diseases such as SLE. Most autoantibodies characterized to date are directed against isolated nuclear molecules such as DNA or histones. We have obtained from spontaneously autoimmune mice six IgG mAb that recognize conformational nucleosome epitopes, but do not react with individual histones or DNA. For three of these mAb, the epitope is at least partially present in the H2A-H2B-DNA nucleosome subparticle, although their binding characteristics differ from those of conventional anti-H2A-H2B-DNA antibodies. All six mAb use VH or Vkappa genes which are recurrently utilized in anti-DNA and other antinuclear antibodies. The V regions of the nucleosome-reactive mAb also contain charged (mostly cationic) residues at sites that are likely to be critical for interaction with nucleosomal antigens. These results suggest that the usage of certain V gene segments in conjunction with suitable V(D)J rearrangements may confer reactivity to nucleosomal antigens. B cells producing such autoantibodies are probably expanded early during the autoimmune process. Somatic mutations in the V regions of nucleosome-reactive mAb may modulate their specificities and result in the acquisition of binding patterns restricted to individual chromatin components such as DNA.