抗伯氏舒螺旋菌表面蛋白A抗體與類風濕關節炎之研究

The Study of Antibody to Outer-Surface Protein A of Borrelia burgdorferi and Rheumatoid arthritis

劉瀚文(Han-Wen Liu)

指導教授 ： 蔡嘉哲

中山醫學大學/醫學院/免疫學研究所/碩士(2005年)

https://doi.org/10.6834/CSMU.2005.00076

摘要

目的：
萊姆病致病菌伯氏疏螺旋體（Borrelia burgdorferi），在其感染晚期會經由伯氏疏螺旋體體表蛋白A (Outer surface protein A, OspA)的分子擬態 (molecular mimicry)所引發自體免疫反應，造成長期的關節滑液膜炎。在萊姆病關節炎的病人中，有70%會產生OspA特異性抗體，並且會隨著萊姆病關節炎的嚴重程度逐漸增加其抗體效價。在本研究中，我們選擇伯氏疏螺旋菌OspA特異性抗體作為偵測的目標，我們想了解不同自體風濕免疫及類風濕性關結炎的病人，與伯氏疏螺旋菌體表蛋白A之間的相關性。亦希望可由這些疾病之進一步分類中，找出某些特定的特徵，並解釋OspA特異性抗體在其中的意義。

材料和方法：
利用molecular cloning的技術將伯氏疏螺旋體中的螺旋體體表蛋白A大量複製並使用Ni-NTA親合性管柱純化蛋白。進一步利用酵素免疫分析法 (ELISA)分析病人血清內含有的抗OspA抗體。收集血清包括類風濕性關節炎（Rheumatoid arthritis, RA）68位、全身性紅斑性狼瘡（Systemic lupus erythematosus, SLE）病人68位、僵直性脊椎炎病人（Ankylosing spondylitis, AS）共12位和乾燥症候群（Söjogren’s syndrome,SS）病人68位，以及76位健康捐血者，

結果：
實驗結果顯示，結果發現總共31位病人有抗OspA IgG抗體，37位病人有抗OspA IgM抗體。其中12位僵直性脊椎炎(AS)病人，有3位(25%)帶有抗OspA IgG抗體，有1位(8.3%)帶有抗OspA IgM抗體。68位SLE病人有7位(10.3%)帶有抗OspA IgG抗體，8(11.8%)位帶有抗OspA IgM抗體。68位SS病人有6位(8.8%)帶有抗OspA IgG抗體，8 (11.8%)位帶有抗OspA IgG抗體。而68位類風濕性關節炎患者(RA)，有15位(22%)帶有抗OspA IgG抗體，20位(29.4%)帶有抗OspA IgM抗體。在RA病人群中，抗OspA IgG抗體相對效價 (titer)顯著與正常族群明顯較高 (p<0.01)，而抗OspA IgM抗體相對效價更是顯著 (p<0.001)。其中，所有帶有OspA特異性抗體的RA病人皆未符合萊姆病血清學診斷標準。

結論：
我們首次在自體免疫病人中偵測到OspA特異性抗體的存在，並且發現類風濕性關節炎的病人族群中帶有之數量及相對效價最高。我們認為在這些自體免疫病人中的OspA特異性抗體是種自體抗體，未來需進一步研究以厘清OspA特異性抗體與類風濕性關節炎之間的關係。

關鍵字

伯氏疏螺旋體 ； 萊姆病 ； 類風濕性關節炎 ； 自體抗體 ； 伯氏疏螺旋體體表蛋白A

並列摘要

OBJECTIVE:
Borrelia burgdorferi (Bb) is the causative agent of Lyme disease. In the chronic stage of Lyme disease, the Outer surface protein A (OspA) of Bb can trigger autoimmunity in the joint by molecular mimicry. IgG antibody responsed to OspA were found in about 70% of patients with Lyme arthritis. We use recombinant protein to detect anti-OspA antibody in patients with autoimmune disease (AID). The relationship between OspA antibody and patients with autoimmune disease were estimated in the study.

METHODS:
Recombinant antigens of OspA of Bb were tested with patients with AID in enzyme-linked immunosorbent assay (ELISA). The serum sample consisted of 68 patients with Rheumatoid arthritis (RA), Systemic lupus erythmatosus (SLE), Söjogren’s syndrome (SS), and 12 patients with Ankylosing spondylitis (AS). The 76 serums of health group were also obtained.

RESULT:
In analyses of anti-OspA IgG antibodies, 12(22%) of 68 patient serums with RA, 7(10%) of 68 patient serums with SEL, 6(8.8%) of 68 patient serums with SS and 3(25%) of patient serums with AS showed reactivity to OspA. In analyses of anti-OspA IgM antibodies, 20(29.4%) of 68 patient serums with RA, 8(11.8%) of patient serums with SLE, 8(11.8%) of patient serums with SS and 1(8.3%) of 12patient serums with AS, showe reactivity to OspA. In patient group of RA, the titer of anti-OspA IgG antibodies was significantly higher than normal group (p<0.05). The titer of anti-OspA IgM antibodies was more significant (p<0.001). The serologic tests for diagnosis of lyme disease have no response in all of the patients with anti-OspA antibodies

CONCLUSION:
This is first time that detect anti-OspA antibody in AID patients. Our result shows that the patients with RA were higher anti-OspA antibody and the most in AID patient. We suggest that the anti-OspA antibody was an autoantibody. The relationship between OspA antibody and rheumatoid arthritis should be estimated in the future.

並列關鍵字

Borrelia burgdorferi ； Lyme disease ； Rheumatoid arthritis ； autoantibdoy ； OspA

參考文獻

1. Steere, A.C., Lyme disease. N Engl J Med, 1989. 321(9): p. 586-96.

2. Kurtenbach, K., D. Carey, A.N. Hoodless, P.A. Nuttall, and S.E. Randolph, Competence of pheasants as reservoirs for Lyme disease spirochetes. J Med Entomol, 1998. 35(1): p. 77-81.

3. Steere, A.C., S.E. Malawista, D.R. Snydman, R.E. Shope, W.A. Andiman, M.R. Ross, and F.M. Steele, Lyme arthritis: an epidemic of oligoarticular arthritis in children and adults in three connecticut communities. Arthritis Rheum, 1977. 20(1): p. 7-17.

4. Burgdorfer, W., A.G. Barbour, S.F. Hayes, J.L. Benach, E. Grunwaldt, and J.P. Davis, Lyme disease-a tick-borne spirochetosis? Science, 1982. 216(4552): p. 1317-9.

5. Sigal, L.H., Lyme disease: a review of aspects of its immunology and immunopathogenesis. Annu Rev Immunol, 1997. 15: p. 63-92.