This study aims to describe a novel antinuclear antibody directed to proteins only accessible during the mitosis: anti-"dividing cell antigen" (DCA) antibody. A total of 709 disease-associated and control sera was tested by indirect immunofluorescence using a variety of cell lines as substrate. Cells were treated with enzymes and antibodies absorbed with nuclear antigens. Antibodies to DNA, histone subfractions, and synthetic peptides were evaluated using enzyme-linked immunosorbent assays. Cell extracts were electrophoresed before and after synchronization and sera tested on the blots. The anti-DCA antibody was demonstrated in 10 of 183 SLE patients but virtually never in other connective tissue diseases. The DCA was sensitive to HCl and proteolytic enzymes and the anti-DCA binding inhibited by histones H2A and H2B. Differences of anti-H2A and anti-H2B were observed between anti-DCA antibody-positive and anti-DCA antibody-negative sera, and antibodies specific for the 1-15 region of H2A, the 1-25 region of H2B and the 1-29 region of H4 were more frequent in the former sera than in the latter. The anti-DCA antibody was shown to react with a 60-kDa protein. Our findings suggest that the anti-DCA antibody is directed to a protein complex containing H2A and H2B.