The use of adenovirus-infected HeLa cells for the detection of low titer autoantibodies.
Following infection of HeLa cells with adenovirus type 5 the cellular La protein becomes predominantly associated with the virally encoded RNA polymerase III products VAI, and VAII, while most of the host RNA polymerase II (e.g. U1, U2, U4, U5 and mRNA) and RNA polymerase III transcription (e.g. U6 and pre-tRNAs) ceases. Other RNA polymerase III products such as the cellular Ro RNAs continue to be transcribed and assembled into ribonucleoprotein complexes containing the Ro (SS-A) antigens. Using a 32P-pulse chase-labeled, adenovirus-infected HeLa cellular extract as a source of antigen, anti-La (SS-B) and anti-Ro (SS-A) antibodies can be detected simultaneously using an immunoprecipitation assay. In the present study this method was found to be more sensitive in detecting anti-La antibodies then counter immunoelectrophoresis and immunoblotting. In studies of sera from patients suffering from rheumatic diseases the percentage positive for anti-La antibody was significantly elevated using this method, especially in patients with systemic lupus erythematosus.
Slobbe
R
R
Department of Biochemistry, University of Nijmegen, The Netherlands.
Van Esch
B
B
Kveder
T
T
Van Venrooij
W J
WJ
D000260:Adenoviruses, Human / Q000276:immunology; D000974:Antibodies, Antinuclear / Q000032:analysis; D001323:Autoantibodies / Q000032:analysis*; D001324:Autoantigens / Q000235:genetics / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells / Q000276:immunology; D006801:Humans; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D011233:Precipitin Tests; D012313:RNA / Q000032:analysis; D020733:RNA, Small Cytoplasmic; D012216:Rheumatic Diseases / Q000276:immunology*; D012261:Ribonucleoproteins / Q000235:genetics; D017411:Ribonucleoproteins, Small Nuclear; D000097585:SS-B Antigen; D000099207:SS-A Antigen
D000974:Antibodies, Antinuclear; D001323:Autoantibodies; D001324:Autoantigens; D012313:RNA; D020733:RNA, Small Cytoplasmic; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear; D000097585:SS-B Antigen; D000099207:SS-A Antigen; C116006:RO60 protein, human; C035356:SS-A antibodies; C035357:SS-B antibodies