2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38039Rheumatoid arthritis (RA) is a common autoimmune disease that affects about 1% of the population in the world. The typical manifestation of RA is chronic, symmetric polyarthritis. As the disease progression, it will cause severe joint destruction and deformities. In the major of the patients with RA, there is a kind of highly specific autoantibodies, which can recognize (or binding with) several citrulline containing proteins, such as filaggrin, fibrin and vimentin, etc. We call such kind of autoantibodies as anti-citrulline-containing protein antibodies. The presence of the citrulline in the protein is caused by deimination by a special enzyme, peptidylarginine deiminase (PAD). The sensitivity and specificity of anti-cyclic-citrullinated peptide antibody (anti-CCP antibody, one of anti-citrulline-containing protein antibodies) are about 70% and 96%, respectively (Schellekens GA, et al. 1998; Schellekens GA, et al. 2000). In previous studies, there are many plasma cells which producing anti-CCP antibodies appearing in the inflamed synovial membrane of RA. These findings provide evidences for an antigen-driven maturation of CCP-specific B cells at the site of inflammation in RA (Reparon-Schuijt,-C-C, et al. 2001). In the meantime, deiminated fibrin α and β chains are the major targets in the inflamed synovial membrane of the patients with RA (Masson-Bessiere,-C, et al. 2001). PADI type 4 distributes in the granulocytes and monocytes (Erik R. Vossenaar, et al. 2003). In Japan, a case control study showed that PADI4 haplotype associated with susceptibility to RA increases production of deiminated peptides acting as autoantigens (Akari Suzuki et al. 2003). Therefore, protein deimination by PADI IV may play an important role in autoantigen formation, autoantibody generation and the pathogenesis of RA. We focus on protein deimination here, and hope these works will contribute to the pathogenesis, clinical diagnosis and treatment of RA. Although the importance of PADI IV is clear, its regulation is still unknown. In addition, protein deimination occurs during the process of apoptosis. During the apoptosis, vimentin is deiminated, and deiminated vimentin could not be polymerized (Asaga H, et al. 1998; Mizoguchi M, et al. 1998). The findings indicate vimentin deimination is associated with the morphologic change of apoptosis. In the granulocytes, PADI IV deiminates nuclear histones during apoptosis. Deiminated histones become less positive charged and unfolded. Both the conformation and the net charge of the histones are altered by deimination, potentially causing the nucleosomes to open up, which makes the DNA more susceptible to degradation by DNase. We could predict PADI IV also plays an important role in DNA fragmentation of apoptosis. The regulation and the role of PADI IV in apoptosis are other purposes of this aim. We divide the program into six aims in three years as following: The first year: 1. The Sensitivity and Specificity of Anti-Citrulline-Containing Proteins Antibodies in the Patients of Rheumatoid Arthritis in Taiwan. 2. The Association between Single Nucleotide Polymorphism in PADI IV, HLA DR4 Gene Complex and Rheumatoid Arthritis in Taiwan. The second year: 3. New Citrulline-Containing Auto-antigens Extracted from the Synovial Membrane of the Patients with Rheumatoid Arthritis, or by Deimination of Proteins Extracted from HL-60 Cells, the Synovial Membrane of Patients with Rheumatoid Arthritis and Osteoarthritis. 4. The Epitope of New Citrulline-Containing Auto-antigen of Rheumatoid Arthritis. The third year: 5. The Functional Differences and the Importance in the Generation of Citrulline-Containing Auto-antigens about Different Single Nucleotide Polymorphisms in PADI IV Enzyme Associated with Rheumatoid Arthritis. 6. The Regulation of PADI IV and the Role of PADI IV in Apoptosis.類風濕性關節炎是最常見的自體免疫疾病，全球人口大約有百分之一的人罹 患此病。典型類風濕關節炎的病程為慢性對稱性多發性關節發炎。疾病持續地進 展下去會導致嚴重的、無法挽回的關節破壞及變形。 在多數類風濕性關節炎病患血清中存在一類具有高專一性的抗體。這類抗體 的特色是會辨識一些含瓜胺酸蛋白質(citrulline containing proteins)如filaggrin, fibrin, vimentin，因此稱為抗含瓜胺酸蛋白質抗體(anti-citrulline-containing protein antibodies)。瓜胺酸所以在蛋白質上出現是經由一種稱為胜肽精胺酸脫亞氨酶 (peptidylarginine deiminase, PADI)的酵素將蛋白質上的精胺酸(arginine)轉成瓜胺 酸(citrulline) 。將一段特殊的含有瓜胺酸的氨基酸片段當成抗原， 運用 enzyme-linked immunosorbent assay來檢測類風濕關節炎病患，其敏感性可達 70%，而專一性更高達96% (Schellekens GA, et al. 1998; Schellekens GA, et al. 2000)。後續的研究發現在病人發炎的關節滑液膜中存有產生抗含瓜胺酸蛋白質 抗體的漿細胞(plasma cells)，因此有可能在滑液膜中存在著含瓜胺酸蛋白質，刺 激B細胞的分化與抗體的產生(Reparon-Schuijt,-C-C, et al. 2001)。同時關節滑液膜 中脫亞氨化的fibrin α鏈與β鏈 (the α chain and β chain of fibrin)被認為主要的自體 免疫抗原(Masson-Bessiere,-C, et al. 2001)。第四型胜肽精胺酸脫亞氨酶 (PADI IV) 分布於免疫細胞的顆粒球(granulocyte)及單核球(monocyte) (Erik R. Vossenaar, et al. 2003)。日本發表的一篇大型case-control研究，發現PADI IV基因上的單一核苷 酸多型性(single nucleotide polymorphism)許多位點與類風濕性關節炎有極大的相 關性(Akari Suzuki et al. 2003)。 因此第四型胜肽精胺酸脫亞氨酶所造成的蛋白質脫亞氨化在自體抗原的形 成、自體抗體的產生及致病機轉上扮演重要角色。在這個子計畫中我們將目標集 中在這個方向，期望能對於類風濕性關節炎致病機轉、臨床診斷及治療上有所貢 獻。 另外第四型胜肽精胺酸脫亞氨酶的調節目前還不清楚。在有限的基礎研究文 獻上顯示它在細胞凋亡過程中可能扮演一些角色。蛋白質的脫亞氨化發生在細胞 凋亡的過程中，特別是細胞骨架中的vimentin被脫亞氨後便無法在聚合，這可能 與凋亡細胞的型態變化有關(Asaga H, et al. 1998; Mizoguchi M, et al. 1998) 。在顆 粒球中第四型胜肽精胺酸脫亞氨酶會讓細胞核內的histone脫亞氨化，使得histone 的正電荷減少並改變蛋白質的結構，進而使histone無法與DNA緊密結合。這被認 為與細胞凋亡中的DNA片段化(DNA fragmentation)有關。所以第四型胜肽精胺酸 脫亞氨酶的調節及它與細胞凋亡之間的關係也是我們研究的重點。 在這個部份的三年子計畫分為六個主題，分列如下： 1. 台灣類風濕關節炎病患對於抗含瓜胺酸蛋白質抗體檢驗的敏感性與特異性研 究。 The Sensitivity and Specificity of Anti-Citrulline-Containing Proteins Antibodies in the Patients of Rheumatoid Arthritis in Taiwan. 2. 台灣類風濕關節炎病患與人類淋巴球抗原DR4和第四型胜肽精胺酸脫亞氨酶 基因上的單一核苷酸多型性相關性分析。 The Association between Single Nucleotide Polymorphism in PADI IV, HLA DR4 Gene Complex and Rheumatoid Arthritis in Taiwan. 3. 蛋白質脫亞氨化修飾後的抗原研究：發現新抗含瓜胺酸蛋白質抗原 New Citrulline-Containing Auto-antigens Extracted from the Synovial Membrane of the Patients with Rheumatoid Arthritis, or by Deimination of Proteins Extracted from HL-60 Cells, the Synovial Membrane of Patients with Rheumatoid Arthritis and Osteoarthritis. 4. 新抗含瓜胺酸蛋白質抗原之免疫抗原決定位點研究 The Epitope of New Citrulline-Containing Auto-antigen of Rheumatoid Arthritis. 5. 不同單一核苷酸多型性基因的第四型胜肽精胺酸脫亞氨酶之功能差異與產生 免疫抗原決定位點之重要性研究 The Functional Differences and the Importance in the Generation of Citrulline-Containing Auto-antigens about Different Single Nucleotide Polymorphisms in PADI IV Enzyme Associated with Rheumatoid Arthritis. 6. 第四型胜肽精胺酸脫亞氨酶之調節與在細胞凋亡中之角色 The Regulation of PADI IV and the Role of PADI IV in Apoptosis.Project