| Summary: | 碩士 === 中山醫學院 === 免疫學研究所 === 89 === Thirty three sera samples from anticentromere antibody(ACA) positive patients identified by ANA(anti-nuclear antibody) screen were used in the present study. In order to characterize the binding sites of autoimmune antibodies in human and other mammalian chromosomes, human HEp-2 and Indian Muntjac metaphase chromosomes were used as substrate for indirect immunofluorescence investigation. By taking the advantage of large Indian Muntjac centromeres , attempts were made to find the difference of antibodies in different autoimmune diseases. These two substrates were also used in Western blot experiments, to analysis the autoantigens recognized by the serum. The ACA positive patients were found to have one or more following syndromes ; Including Raynaund’s syndrome , Rheumatoid Arthritis, Diabetes Mellitus, Sjogren’s syndrome, CERST syndrome, REST(incomplete CREST), Primary Biliary Cirrhosis(PBC). In the sera of ACA positive patients, two were found to have fluorescence signals not only in kinetochore/centromere domain but also in whole chromosome. This finding is similar to those Systemic Lupus Erythematosus (SLE) patients reported by Martin et al (1990) [22][fig12 ]. Therefore several rheumatic disease patients serum , such as SLE, Rheumatoid Arthritis(RA), Ankylosing spondylitis(AS) were chosen for further study. Such study involved the comparison of immunofluorescence differences between different rheumatic diseases, and to search the specific marker between different diseases. Five fluorescence patterns were found .Negative, KD(kinetochore domain), WC(whole chromosome arms), KD＋WC, CD(centromere domain)＋WC individual. However ACA positive patients were found to present mostly with KD pattern only；SLE patients present with KD＋WC pattern in the majority of the cases，and most RA patients were found to be immunofluorescence negative. According to the results, Indian muntjac chromosome has highly sensitivity to analysis the immunofluorescence pattern of autoantibodies. So I recommend that Indian muntjac chromosome can be used in clinical ANA screen.
In antigens study, 30 patients sera recognized Indian Muntjac CENP-A (93.9%), 29 patients sera recognized HEp-2 CENP-A (87.8%), 33 patients sera recognized Indian Muntjac CENP-B (100%), 31 patients sera recognized HEp-2 CENP-B (93.9%), 31 patients sera recognized Indian Muntjac and HEp-2 CENP-B (93.9%). The results exhibite that anticentromere antibodies doesn’t specifically presented in a particular autoimmune disease, and no specific serum against specific antigen was found.
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