| 摘要: | 首先從抗核抗體(anti-nuclear antibody)篩檢鑑定過的血清檢體中，挑選33例判別為抗染色體著絲點抗體陽性的血清檢體，利用間接免疫螢光染色法，並使用人類喉癌上皮細胞(HEp-2)及印度山羌(Indian muntjac)細胞的有絲分裂中期染色體當受質，試圖來比較自體抗體在人類及其它哺乳類著絲點上結合的情形，並利用Indian muntjac分裂中期染色體具有大且易觀察的著絲點，找出不同自體免疫疾病之間所具有的抗體差異性。同時使用此兩種受質進行西方點墨法來分析這些血清認識的自體抗原。在抗著絲點抗體陽性病人中，都會有一個或更多的下列症狀被發現，包括雷諾氏症候群、類風濕性關節炎、糖尿病、修格連氏症候群、CREST症候群、REST(incomplete CREST)、原發性膽汁性肝硬化。在抗染色體著絲點抗體陽性血清中，發現到2位病人除了在著絲點的部位有螢光反應外，在染色體其它的部分也有反應，這個結果和Martin et al.(1990)[22]所提出紅斑性狼瘡病人的螢光染色有相似之處，因此選擇幾種免疫風濕性的疾病，如紅斑性狼瘡、風濕性關節炎、僵直性脊椎炎的病人血清，來進一步比較不同疾病之間螢光反應的差別，並試圖找到不同疾病的專一性標記。結果發現可以分為5種不同的螢光表現型，分別為陰性(negative)、KD(kinetochore domain) only、WC(whole chromosome arms) only、KD＋WC、CD(centromere domain)＋WC；而抗著絲點抗體陽性病人大部分出現KD only pattern，紅斑性狼瘡病人大部分出現KD＋WC pattern，風濕性關節炎病人大部分為陰性，僵直性脊椎炎病人大部分出現KD only pattern。由以上之結果可知，Indian muntjac染色體對於研究自體抗體的螢光表現型具有相當高的靈敏度，故建議臨床上可改用Indian muntjac染色體來進行抗核抗體篩檢。
在抗原分析方面，33位病人的血清中，有30位血清認識Indian muntjac CENP-A(17KD)占93.9%、29位血清認識HEp-2 CENP-A占87.8%；33位血清認識Indian muntjac CENP-B占100%、31位血清認識HEp-2 CENP-B占93.9%；31位血清認識Indian muntjac 及HEp-2 CENP-C占93.9%。結果顯示抗著絲點抗體並不只出現於特定自體免疫疾病，而且並無特定疾病血清對抗特定抗原的發現，這個結果與之前其他作者發表的結果相似。
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. |
