2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38546The mechanism of autoimmune disease remains unclear. However, it has been thought that balance of cell survival and death is the main reason of autoimmune disease. Recent research showed that apoptotic cells serve as the main repertoire of autoantigens. Moreover, the deficiency of cells to recognize and clear apoptotic body is a main factor of the induction of autoimmune responses. Therefore, it becomes important to study the mechanism involved in the production, recognition and clearance of autoantigens. Transglutaminase 2 is a multifunctional enzyme with the function of cross-linking Ca2+-dependent reactions which result in post-translational modification of substrate proteins. Results from several independent laboratories indicated that transglutaminase 2 plays a potential role in cell apoptosis. From these results, we suppose that transglutaminase 2 may be involved in the process of neo-autoantigen production after cell apoptosis and also the clearance of apoptotic body. In this study, we aim to investigate the factors (including TGF-β and IL-10) of affecting phagocytosis and clearance of apoptotic cells in autoimmune diseases. In the first year, UV irradiation is used to induce karatinocytes apoptosis in order to establish the suitable material to assay phagocytosis of macrophage. To further define the role of TG2 in phagocytosis, we will examine TG2 expression in apoptotic cell, phagocytes and their effects on the uptake of apoptotic cell. As clearance of apoptotic cell is regulated by multiple factors, in the second year, we will focus on relationship between the regulation factors such as cytokines of TGF-β or IL-10 and TG2 in process of phagocytosis. Besides, we will use the TG2 inhibitor like Cystamine and TG2 RNAi to monitor its role in the process of phagocytosis. Finally, we will study the role of TG2, TGF-β and IL-10 in phagocytosis of monocyte and PMN from patients with SLE. The first year: To establish ultraviolet irradiation system to induce apoptosis of keratinocytes in order to assay phagocytosis of macrophage. The second year: To investigate the relationship between regulation factors like cytokines of TGF-β or IL-10 and TG2 in process of phagocytosis by TG2 RNAi and TG2 inhibitor. The third year: The macrophage and PMN or multiple regulation factors derived from patients with SLE and normal controls will be used to monitor the expression of TG2 in the process of clearance. The study of apoptosis and clearance of apoptotic cells may help us more understand its pathogenesis in autoimmune diseases, and have some contribution for its further treatment in the future.自體免疫疾病的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一直被認為是造成自體免疫疾病的主要原因，近年來的研究更指出，細胞凋亡的過程中會產生自體抗原，而凋亡小體無法被正確的辨識及清除，則會產生自體免疫反應，因此，研究自體抗原的產生以及清除，將是自體免疫疾病產生之決定性關鍵。Transglutaminase 2(TG2)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修飾，最近幾個獨立實驗室的研究發現，TG2是參與細胞凋亡的一個重要潛在成員，我們因此懷疑TG2及其他cytokines如IL-10及TGF-β是否參與細胞凋亡，甚至影響巨噬細胞辨識及清除凋亡小體，進而驅動自體免疫反應的過程。本研究主要在於分析影響吞噬作用及清除凋亡細胞的因子。我們首先建立細胞凋亡及吞噬作用的模式，利用UV照射人類角質細胞株製備細胞凋亡小體，以作為研究巨噬細胞細胞株吞噬作用的材料，並觀察細胞受到UV作用下TG2的變化，進一步探討TG2與吞噬作用過程的關係，我們同時也觀察TGF-β及各種相關的細胞激素如IL-10、TGF-β對TG2和吞噬過程的影響，除此之外，也將利用RNAi技術和TG2酵素抑制劑(Cystamine)，以釐清TG2的角色。最後我們將持續收集臨床自體免疫病患檢體，包括病人血清，以及分離血球如monocyte、PMN，比較相關因子在正常人及病患體內的表現情形是如何，以及SLE病人吞噬能力會受到哪些外在因素的影響，以探討在臨床病人體內，調節自體抗原產生以及清除的機制。相信這些相關之細胞凋亡及吞噬研究，將有助於我們更進一步瞭解對自體免疫疾病之調控機制情形。 第一年: 以UV照射建立角質細胞株(HaCaT cell line)凋亡細胞模式以作為巨噬細胞吞噬作用的材料，並瞭解UV對TG2及其他cytokines，如IL-10、TGF-β之變化。 第二年: 在細胞中探討TG2，TGF-β及各種相關的cytokines,IL-10,影響吞噬作用及清除的角色，並利用RNAi技術，探討TG2與吞噬作用的關係。 第三年: 進而探討TG2，TGF-β及各種cytokines,IL-10,在人類SLE中吞噬及清除作用的影響。 結論則是將來在經過這三年的研究，希望能更深入瞭解自體免疫疾病的致病機制及其影響因素，以提供臨床醫療上對診斷及治療有更大的貢獻。autoimmunetransglutaminase 2post-translational modificationphagocytosisProject