TRANSFECTION AND OVEREXPRESSION OF THE HUMAN 60-KDA RO/SS-A AUTOANTIGEN IN HEP-2 CELLS

被引:52
作者
KEECH, CL
MCCLUSKEY, J
GORDON, TP
机构
[1] Centre for Transfusion Medicine and Immunology, Department of Clinical Immunology, Flinders Medical Centre, Bedford Park
来源
CLINICAL IMMUNOLOGY AND IMMUNOPATHOLOGY | 1994年 / 73卷 / 01期
关键词
D O I
10.1006/clin.1994.1181
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
A full-length cDNA encoding human 60-kDa Ro/SS-A protein was transfected into and overexpressed in the human cell line HEp-2, with the aim of developing an improved reagent for indirect immunofluorescence (IF) detection of anti-Ro autoantibodies. Stable transfectants were analyzed by IF using a panel of 20 precipitin-positive anti-Ro human sera. Transfectants showed bright finely speckled nuclear and nucleolar staining. No surface membrane expression was detected despite marked overexpression of 60-kDa Ro. All human anti-Ro sera reacted with the transfectants with titers ranging from 1:320 to 1:40,960. The same sera tested on untransfected cells showed titers from negative (3 sera) to 1:160. These transfectants dramatically increase the sensitivity of IF testing (mean increase in titer of 41-fold) and allow detection of specific anti-Ro antibodies in samples negative or equivocal on untransfected cells. The staining patterns of antisera of other important specificities remained unaltered. In a study of sera from 22 patients with systemic lupus erythematosus, anti-60-kDa Ro autoantibodies were detected in all sera when tested on 60-kDa Ro-transfected HEp-2 cells; however, in 12 of 22 sera autoantibodies were undetectable by recombinant 60-kDa Ro ELISA. The 60-kDa Ro transfectants are a simple and sensitive method for detection of anti-Ro antibodies in patients with systemic rheumatic diseases. (C) 1994 Academic Press, Inc.
引用
收藏
页码:146 / 151
页数:6
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