Studies on Antibodies to Histones by Immunofluorescence

When mouse kidney tissue sections were extracted with 0.1 N hydrochloric acid, sera with antibodies to certain nuclear antigens no longer stained tissue nuclei by immunofluorescence. This effect was due to removal of histones and nuclear acidic proteins Sm and nuclear ribonucleoprotein by the acid. DNA remained in the nuclei of the acid-extracted tissue sections. When solutions of calf thymus histones were reacted with acid-extracted tissues, histones combined with nuclear DNA to form complexes of DNA-histone. These complexes contained antigenic determinants which reacted with sera containing antibodies to deoxyribonucleoprotein to give nuclear staining demonstrated by immunofluorescence. The reaction was immunologically specific in that sera with antibodies to Sm and nuclear ribonucleoprotein were not reactive with reconstituted DNA-histone in nuclei. Other basic proteins such as protamine, poly-L-lysine, and poly-L-arginine could not substitute for histones. The method is introduced as a specific and reproducible assay for study of antibodies to histones.