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  • 學位論文

類風濕性關節炎與抗牙齦痛蛋白A之抗體

Rheumatoid Arthritis and Anti-Arg-Gingipain A Antibody

指導教授 : 蔡嘉哲

摘要


目的: 類風溼性關節炎(RA)與牙周病(PD)關係密切,兩者的病徵很類似 都是慢性發炎造成關節或是齒槽骨的損傷。有研究指出類風濕性關節 炎的病患罹患牙周病的比率比起其他人約高出兩倍。造成牙周病的主 要致病菌牙齦卟啉單孢菌(Porphyromonas gingivalis)有許多毒理因子 不但可以引起發炎反應,甚至可以幫助細菌躲避免疫系統的攻擊,其 中一重要的毒理因子: Arg-gingipain A (RgpA)已經被證實可以分解宿 主的免疫球蛋白、補體、以及細胞激素等,並且會造成血管通透性增 加進而加劇發炎反應。本研究想藉由偵測類風濕性關節炎以及其他自 體免疫病患血清中是否有抗RgpA抗體以及觀察病情嚴重程度是否與 抗體效價有關連。 材料與方法: 利用molecular cloning 技術將Porphyromonas gingivalis 之RgpA 蛋白分成酵素活性區(catalytic domain)以及血球凝集區(hemagglutinin domain)分別大量表達後純化,再以酵素免疫分析法(ELISA)分析90 位類風溼性關節炎(rheumatoid arthritis, RA)、38 位全身性紅斑性狼瘡 (systemic lupus erythematosus, SLE)、15 位非RA、SLE 之自體免疫病 患以及11 位對照組(normal control)血清內是否含有抗hemagglutinin domain 或抗catalytic domain 之抗體。 結果: 實驗結果發現共有40 位病人有抗RgpA hemagglutinin domain 的 抗體,0 位病人有抗RgpA catalytic domain 抗體。其中90 位RA 病人 中有29 人帶有抗RgpA hemagglutinin domain 抗體(32.2%),38 位SLE 病人中有6 人帶有抗RgpA hemagglutinin domain 抗體(15.8%),15 位 非RA、SLE 之自體免疫病人中有5 人帶有抗RgpA hemagglutinin domain 抗體(33.3%)。在RA 病人群中,抗RgpA hemagglutinin domain 抗體陽性比率明顯高於對照組(p<0.05),而所有族群之抗RgpA catalytic domain 抗體與對照組相比則無統計學上的意義(p>0.05)。 結論: 我們首次偵測及分析自體免疫病人中抗RgpA 蛋白之抗體情形。 結果顯示RgpA catalytic domain 可能較不具抗原性,所以在所有人的 血清中都無法偵測到特異性抗體存在。而在RgpA hemagglutinin domain 方面,RA 病人中帶有RgpA hemagglutinin domain 抗體的陽性 比率明顯高於對照組及其他族群,這也和先前其他團隊研究RA 病人 較容易罹患牙周病結果一致,未來需進一步釐清RgpA 如何參與RA 之致病機轉。

並列摘要


Objective Increasing evidences have suggested a high correlation between rheumatoid arthritis (RA) and periodontitis (PD). Both inflammatory diseases share a common aetiololgy and pathology characterized by joint destruction or alveolar bone resorption. Previous studies have shown that patients with RA have a two times increased incidence of expressing mild to severe PD, compared with healthy people. The main pathogen of periodontitis, Porphyromonas gingivalis, possesses numerous virulence factors which can elicit severe immune response. Arg-gingipain A (RgpA), one of the virulence factors , has been reported to degrade a variety of host proteins, including immunoglobulins, complement proteins, cytokines, and increases vascular permeability to enhance leukocyte recruitment to the infected region, and finally caused inflammation. In this study, we detect anti-RgpA antibody in patients with autoimmune disease by using recombinant RgpA protein and further analyze whether there is a correlation between anti-RgpA antibody and RA. Methods: RgpA hemagglutinin domain and RgpA catalytic domain were cloned respectively. ELISAs using RgpA hemagglutinin and RgpA catalytic domain were developed and antibody reactivities examined in patients with RA (n=90), SLE (n=38), healthy normal control (n=11), and individuals with neither RA nor SLE patients (n=15). Results: In analysis of anti-RgpA catalytic domain antibodies, no one was found to react with RgpA catalytic domain. In analysis of anti-RgpA hemagglutinin domain antibodies, 29 of 90 RA patients (32.2%), 6 of 38 SLE patients (15.8%), 5 of 15 neither RA nor SLE patients (33.3%) were found to react with RgpA hemagglutinin domain. The titer of anti-RgpA hemagglutinin domain antibodies in RA patients was significantly higher than that of the healthy normal control group (p<0.05). Antibodies to RgpA catalytic domain in each of disease groups were found no significant difference compared to that of healthy normal control group (p >0.05). Conclusion: Our results showed that RgpA catalytic domain does not possess antigenic property whereas RgpA hemagglutinin domain exhibit excellent antigenic property which can elicit immune response to produce specific antibodies against themselves. Patients with RA have higher titer of anti-RgpA hemagglutinin domain antiboies among the disease groups. Our results were also consistent with other people’s study that patients with RA had a two times increased prevalence of expressing mild to severe PD. The relationships between anti- RgpA hemagglutinin domain antiboies and rheumatoid arthritis need to be further elucidated.

並列關鍵字

Rheumatoid arthritis Periodontitis

參考文獻


Molecular cloning and structural characterization of the Arg-gingipain
15. O'Brien-Simpson NM, Black CL, Bhogal PS, Cleal SM, Slakeski N, Higgins
Herwald H, Nguyen KA, Eick S, Potempa J, Kozik A: Adsorption of
34. Whitney C, Ant J, Moncla B, Johnson B, Page RC, Engel D: Serum
1. McInnes IB, Schett G: Cytokines in the pathogenesis of rheumatoid

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