Human autoantibodies that react with both cell nuclei and plasma membranes display specificity for the octamer of histones H2A, H2B, H3, and H4 in high salt

Sera of some patients with systemic lupus erythematosus and related diseases contain a polyclonal antibody population (cross-reactive antinuclear antibodies [X-ANA]) that react specifically with both core mononucleosomes and plasma membranes of viable nucleated cells. Native mononucleosomes and nucleosome cores assembled from long DNA and the inner histones were indistinguishable in terms of inhibition of binding of X-ANA to nuclei of tissue sections and to polynucleosomes on the walls of plastic tubes. In contrast, mononucleosomes selectively depleted of histones H2A and H2B did not inhibit these reactions. A method was developed for isolation of X-ANA from serum that took advantage of the dual specificity of these antibodies. Immunosedimentation in sucrose density gradients revealed that 125I-labeled Fab' fragments of highly pure X-ANA formed complexes with the inner histones H2A, H2B, H3, and H4 in 2 M NaCL, but not in 0.15 M salt. These results indicate that X-ANA recognize an epitope of the inner histone in 2 M salt, and that in 0.15 M NaCL this epitope is not formed unless the histones interact with DNA to generate a nucleosome structure. Furthermore, in light of the previous demonstration that the epitope is destroyed by trypsin, it may be localized in the N-terminal region of histone H2A or H2B.