2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38037The mechanism of autoimmune diseases is still unclear. But the cell survival and death balance has been considered and is being as the main reason of autoimmune diseases. Apoptosis or programmed cell death (PCD) is an active process of cellular self-destruction, essential for embryonic development and maintenance of homeostasis of multicellular organisms. Programmed cell death induction can serve as a defence mechanism of the host against intracellular microbes. Virus infections trigger host cell apoptosis, which can either limit virus production or contribute directly to viral pathogenesis. Several independent laboratories have identified transglutaminase 2 (TG2) as a potentially important player of the cell death program(s). This gene is specifically expressed in cells dying during mammalian development as well as in those undergoing apoptosis in various patho-physiological and experimental settings. The transglutaminase 2 is a multifunctional enzyme in its cross-linking configuration catalyzes Ca2+ -dependent reactions resulting in post-translational modification of proteins by establishing epsilon (gamma-glutamyl) lysine cross-links and/or covalent incorporation of biogenic amines (di- and poly-amines and histamine) into proteins. In autoimmune diseases including systemic lupus erythematosus (SLE), the immune system attacks various autoantigens and causes damage in target organs. Recently, it was found that dead cells serve as a repertoire for autoantigens which can stimulate an autoimmune response in sensitive persons. The mechanisms which lead to induction and progress of apoptosis include extra-cellular stimuli, intra-cellular signals, and cleavage of proteins. During apoptosis, several events occur including migration of intra-cellular components to the cell membrane, removal of apoptotic cells by specific proteins and complement systems, and phagocytosis of apoptotic cells by macrophages. In recent years, evidence has accumulated that the autoimmune response can be initiated as a result of defects in apoptosis and/or removal of apoptotic material. So, in our study, we will study the effect of apoptosis on autoimmune diseases, especially in SLE. In the first year, we will establish and use real-time PCR to study the TG 2 gene expression of lymphocytes, PMN, and erythrocytes derived from patients with SLE and normal controls. Also, we will study the expression of TG 2 of animal model MRL-lpr/lpr including its heart, kidney, lung, and spleen etc. As its expression is regulated by multiple factors, in the second year, we will focused on the promoter region of TG2 gene and the polymorphism, and other factors like cytokines of TGF-βand the signal effect of Ras, Raf, and MEK1/ERK. Besides, we will study the role of TG 2 in phagocytosis of monocyte and PMN from patiensts with SLE. In the last year, we will collect apoptotic blebs from B19-NS1 transfected cells and used it as antigens to immunizated Balb/c mice since it is established in our laboratory and known that huamn parvovirus B19-NS1 can induce apoptosis and finding the expression of new antigens. And, we will also use H2O2 to induce apoptosis and immunizated Balb/c mice. These study of apoptosis and clinics may help us more understand its regulation in autoimmune diseases.自體免疫疾病之致病機轉到目前為止仍然不是很清楚，但長久以來細胞存活和死 亡的平衡失調則一直被認為是造成自體免疫疾病之主要原因。細胞凋亡或細胞程 序死亡(PCD)是細胞主動的自毀行動,重要性在於能衡定胚胎發展和個體生長維 護。細胞凋亡之產生可擔當主人的防禦機制對抗細胞內微生物感染生長。病毒感 染宿主細胞而使細胞產生凋亡,這不但限制了病毒生長亦直接地對病毒發生作 用。最近幾個獨立實驗室辨認了transglutaminase 2 (TG2)是為參與細胞死亡的一 個潛在地重要成員，不僅如此，這個基因亦被證實在各種各樣的細胞死亡病理生 理實驗皆有重要的影響。所謂TG 2 是一種多功能酵素，能連結催化仰賴鈣離子 的反應而造成蛋白質轉譯後的改變(藉由建立epsilon(gamma-glutamyl) lysine 連 結和/或共價形成biogenic amines (di- and poly-amines and histamine). 而在自體免疫的疾病中包括全身性紅班性狼瘡(SLE),已知其免疫系統會攻擊各 樣的自體抗原而造成損傷目標器官。最近,它被發現其死亡細胞會起保留作用成 為一個自體抗原因而引發自體免疫反應產生。這死亡機轉進展包括細胞外刺激的 機制,細胞內信號,和蛋白質切割。在細胞凋亡期間,包括細胞內成分對細胞膜的遷 移, 專一蛋白質和補體系統撤除死亡細胞, 及由巨噬細胞吞噬死亡細胞。近年來, 證據積累發現自體免疫反應可能被誘發於瑕疵之細胞凋亡或細胞凋亡之材料撤 除上。因此在我們的研究中，我們將探討自體免疫疾病病人細胞凋亡對疾病之種 種影響，特別是全身性紅班性狼瘡。第一年將建立及利用Real-time PCR 來了解 全身性紅斑性狼瘡病人和正常人其Transglutaminase2 於血液細胞(lymphocytes, PMN 和erythrocytes)表現差異情形。同時亦探討動物模式之紅斑性狼瘡老鼠其各 組織器官包括心、腎、肺、脾等TG 2 之表現。了解其差異後，第二年將更進一 步了解其TG2 promoter 調控之情形,進行polymorphism 研究，和受其他細胞激素 像TGF-β以及Ras、Raf，MEK1/ERK 等分子之影響。另外更進一步也探討病人 之TG2 對於吞噬作用參與之影響，包括參與之吞噬細胞單核球及嗜中性白血球 等。第三年除了延續第一、二年之研究外，我們也將利用我門近幾年來對人類微 小病毒B19 之NS1 蛋白質研究發現其會造成對細胞凋亡現象產生之apoptotic bodies，利用免疫動物探討是否產生或誘發新抗原，以更進一步了解在感染B19 病毒後，引發病人產生自體抗體而造成自體免疫疾病之致病機轉。另外也利用 H2O2 造成細胞凋亡隻抗原探討免疫動物探討是否產生或誘發新抗原，並比較異 同。相信這些相關之細胞凋亡研究，將有助於我們更進一步了解對自體免疫疾病 之調控情形。Systemic lupus erythematosus (SLE) 之Transglutaminase 2 (TG2) 蛋白質表現對Apoptosis 之影響THE EFFECT OF TRANSGLUTAMINASE 2 (TG2) ON APOPTOSIS INSYSTEMIC LUPUS ERYTHEMATOSUSProject