Monoclonal anti-double-stranded deoxyribonucleic antibodies: II. Indirect immunofluorescent studies

Abstract Ten monoclonal anti-deoxyribonucleic acid antibodies (DNA) were generated in three separated fusion experiments using nonimmunized (NZB × NZW)F 1 mouse spleen cells. The specificity of these antibodies was shown to be directed against double-stranded DNA (dsDNA) by radioimmunoassay techniques. The patterns given by these monoclonal anti-dsDNA antibodies in immunofluorescent assay using four different substrates were studied. Most antibodies showed an identical pattern with the same substrate and a different staining with another substrate. All, however, were positive for kinetoplast staining of Crithidia luciliae . These data suggest that a particular substrate favors a specific immunofluorescence pattern. With mouse liver sections, the staining appeared to be dependent upon the antibody tested and technical variables. Particularly, a change of pattern from peripheral to homogeneous with dilution of two monoclonals suggests that the concentration of the antibody may influence the apparent pattern of nuclear staining. In as much as antibodies used are monoclonal and thus directed against a single determinant, one may conclude that the immunofluorescence pattern is not always an adequate indication of antinuclear antibody specificity.