臺灣博碩士論文加值系統

恩瑞舒(ORENCIA®, Abatacept)，是人類細胞毒殺性T淋巴球抗原-4 (CTLA-4)和人類免疫球蛋白G1 (Immunoglobulin G1, IgG1)恆定區(Fc-portion)的融合蛋白(Fusion protein)，目前已在多國核可應用於臨床治療類風濕性關節炎(Rheumatoid arthritis, RA)。其機制是模仿自然產生的CTLA-4和CD28競爭CD80/CD86的結合，進一步的防止T 淋巴球的活化。然而，目前對於CD80/CD86在結合到CTLA-4時，所傳遞給抗原呈現細胞(Antigen-presenting cells)，如:樹突狀細胞(dendritic cells)，巨噬細胞(Macrophage)和B淋巴球的反向信號(reverse signal)尚未被廣泛地探討。我們想藉由活體內(in vivo)及活體外(in vitro)的研究方法，來探討恩瑞舒是否影響B 淋巴球的功能。首先，在臨床RA病人的檢體中，我們發現恩瑞舒可短暫造成周邊血記憶性B淋巴球所表現的CD80/CD86被遮蔽，同時改變周邊血naïve B細胞和記憶性B細胞的比例；然而，恩瑞舒會影響病人中某些專一性抗體的產量。接著在活體外的實驗中，恩瑞舒可抑制某些條件下B 淋巴球活化後產生的分子。一如預期，恩瑞舒能夠抑制非自體(allogeneic) B淋巴球引發的T細胞增生反應。總結我們的結果，我們發現恩瑞舒除了可藉由和CD80/CD86的交互作用，來抑制T淋巴球的反應，另外也可以調控B淋巴球的功能。

Abatacept is a cytotoxic T lymphocyte antigen-4 (CTLA-4) fusion protein approved for rheumatoid arthritis (RA) treatment worldwide. Abatacept is able to mimic the natural CTLA-4 and compete CD28 binding their ligands CD80/CD86 to prevent T cell activation. However, the reverse signals induced through the interaction of CTLA-4-Ig with CD80/CD86 on antigen-presenting cells (APCs) such as dendritic cells (DCs), macrophages and B cells are not fully understood. To test whether CTLA-4 regulates B cell functions, we assayed the effect of Abatacept on human B cells in both in vitro and in vivo conditions. In our study, we observed that Abatacept transiently reduced the level of CD80/CD86 on peripheral blood memory B cells and changed the naïve to memory B cell ratio. We measured several specific antibody levels in the serum from patients with RA before and after Abatacept treatment and observed a differential influence of Abatacept on these antibodies. In the in vitro assays, activation-induced B cell functions were suppressed by Abatacept in certain conditions but not in others. As expected, Abatacept suppressed Daudi B cells-induced allogeneic T cell proliferation, indicating a significant blockade of T-B interaction by Abatacept. Our results demonstrate a profound effect of Abatacept on both in vitro and in vivo B cell activities.

Contents............i
Figure contents............ii
Abstract............iii
1. Introduction............1
1.1 Rheumatoid arthritis (RA)............1
1.2 Abatacept (ORENCIA®)............1
1.3 The co-inhibitory molecule CTLA-4 ............1
1.4 CD80(B7.1) and CD86(B7.2) reverse signaling ............2
1.5 Clinical studies on Abatacept............3
1.6 B cell functions in autoimmune disease............4
1.7 B cell activation ............5
1.8 Hypothesis and specifics aims............5
2. Materials and Methods ............7
2.1 Material............7
2.2 Methods............15
2.3 Statistical analysis............20
3. Results............21
4. Discussion ............28
Acknowledge............33
References ............34
Figures ............39
Supplemental Figures............58

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