| Summary: | 碩士 === 國立陽明大學 === 醫學生物技術研究所 === 87 === Autoantibodies against acidic ribosomal phosphoproteins (anti-P Abs) are detected in 15% of systemic lupus erythematosus (SLE) patients. The presence of the autoantibodies is associated with neuropsychiatric disease, active nephritis, and chronic active hepatitis in lupus patients. To study the pathological role of anti-P Abs in SLE, monoclonal anti-P Abs were established and the effects of anti-P Abs on various cells were investigated. A monoclone 9B6-4 was identified by using standard hybridoma procedure. The antigen specificity of 9B6-4 mAb was demonstrated by ELISA analysis using recombinant human P1 and P2 protein. Western blot also demonstrated that the 9B6-4 mAb recognized P0 (38 kDa), P1 (19 kDa), and P2 (17 kDa). A characteristic P protein in HEp-2 cells was shown by 9B6-4 mAb staining. The 9B6-4 mAb binded to 20—40% of RKO, J774, RBA-1, Jurkat and H1299 cells, and penetrated into 50-90% of these cells as analyzed by indirect immunofluorescence stain and flow cytometry. Further experiments showed that the anti-P mAb penetrated into cells via P0 (38 kDa) surface protein. To investigate the effect of the 9B6-4 mAb penetration into Jurkat T cells, propidium iodide was used to analyze cell cycle. The percentage of sub-G1 cells increased to 14% in cells treated with the mAb. Annexin V-FITC stained 15% of the treated Jurkat T cells which indicated induction of apoptosis..These results suggest that the anti-P autoantibodies may play a role in the pathogenesis of lymphopenia or lymphocyte dysfunction in SLE.
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