2010-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/656867摘要：口腔扁平苔癬（oral lichen planus, OLP）是T細胞免疫異常引起之發炎性疾病。OLP是否為自體免疫疾病，雖然尚未完全確定，但筆者等過去之研究發現，OLP患者有明顯的女性傾向，患者血清中存在多種自體抗體和血中高γ-免疫球蛋白，且有器官特異性和非器官特異性的血清自體抗體的重疊，這些都建議OLP可能是自體免疫疾病。鑑於OLP是T細胞免疫異常引起之發炎性疾病，在過去有針對OLP患者T細胞相關的細胞激素之角色研究，細胞激素干擾免疫調節，導致CD4+細胞中Th1/Th2的失衡，再加上持續的T細胞病理激活，表現出OLP的慢性發炎性質。在很多動物實驗模式，已證明調節性T細胞（Tregs）在自體免疫控制的重要性。此外，有很多研究顯示，人類自體免疫疾病可能存在有Tregs的缺陷。我們初步之研究發現，OLP患者周邊靜脈血液中CD4+/CD8+ 比值和FOXP3+ T 細胞百分比較正常控制組個體為高。OLP患者CD8+ T 細胞產生之IFN-γ 較CD4+ T 細胞產生之IFN-γ為高。因此，本三年研究計畫首先探討有自體免疫傾向之口腔扁平苔癬患者之周邊靜脈血液中CD4+CD25+FOXP3+ Tregs百分比和抑制功能及Th1/Th2細胞激素和TGF-β濃度。使用流式細胞儀，偵測120位OLP患者和60位口腔黏膜健康對照組個體之周邊靜脈血液中CD4+CD25+FOXP3+ Tregs、CD4+CD25+TGF-β+、CD4+CD25+IL-10+之比率。進一步，將每件周邊靜脈血液樣本中之CD4+CD25+ Tregs和CD4+CD25- T細胞分開，觀察CD4+CD25+ Tregs在CD4+CD25- T淋巴細胞複製時的直接抑制作用。在3H-thymidine加入前，收集培養上清液，利用ELISA kit測量IFN-γ, TNF-α, IL-2, IL-4, IL-10和TGF-β的濃度。其次，我們用levamisole治療OLP病人，並追蹤偵測OLP患者接受levamisole免疫調節治療後，其周邊血液中CD4+CD25+FOXP3+ Tregs數量變化及各種細胞激素濃度改變。我們也將使用免疫組織化學染色和RT-PCR，在60例OLP病變處和10例正常口腔黏膜組織中，偵測CD4+CD25+FOXP3+ Tregs數量，並看其和OLP亞型及疾病活性的關聯性。<br> Abstract: Oral lichen planus (OLP) is a T cell-mediated inflammatory oral mucosal disease. It is still not clear whether OLP is an autoimmune disease or not. Our previous studies revealed that OLP patients have a female predilection, have the presence of multiple autoantibodies and high level of γ-immunoglobulin in sera, and have the overlap of presence of both organ-specific and non-organ-specific autoantibodies. These findings suggest that OLP may be an autoimmune disease. As T cell-mediated autoimmunity is considered to be involved in the pathogenesis of OLP, the roles of several T cell-associated cytokines have been investigated in OLP patients in the past decades. Disturbed immune regulation resulting in T helper 1 (Th1)/T helper 2 (Th2) cell imbalance and a continuous pathogenic activation of T cells have been suggested as a cause of chronic inflammation in OLP. The importance of regulatory T cells (Tregs) in the control of autoimmunity is now well established in a variety of experimental animal models. In addition, there are numerous studies suggesting that the Treg deficits may be an underlying cause of human autoimmune disease. Our preliminary studies found that the ratio of CD4+/CD8+ and the percentage of FOXP3+ T cells were higher in OLP patients than in normal control subjects. IFN-γ production by CD8+ T cells was higher than that by CD4+ T cells in OLP patients. Therefore, in this 3-year research project we first investigate the proportion and suppression function of CD4+CD25+FOXP3+ Tregs as well as the levels of Th1/Th2 cytokines and TGF-β in the peripheral blood samples of OLP patients. Flow cytometry is used to detect the proportions of CD4+CD25+FOXP3+ Tregs, CD4+CD25+TGF-β+ and CD4+CD25+IL-10+ cells in the peripheral blood samples of 120 OLP patients and 60 healthy control subjects. Furthermore, CD4+CD25+ Tregs and CD4+CD25- T cells are separated from the peripheral blood sample of each enrolling subject. The direct suppression effect of CD4+CD25+ Tregs on CD4+CD25- T lymphocyte proliferation is performed by mixed lymphocyte culturing. Supernatants are taken from cultures before addition of 3H-thymidine. IFN-γ, TNF-α, IL-2, IL-4, IL-10 and TGF-β levels in supernatants are measured by ELISA kits. Secondly, we will use levamisole to treat OLP patients and evaluate OLP patients’ response as well as changes of the number of CD4+CD25+FOXP3+ Tregs and serum cytokine levels after levamisole immunomodulation therapy. Thirdly, we use immunohistochemistry and real-time RT-PCR to assess the number of CD4+CD25+FOXP3+ Tregs in 60 specimens of OLP lesions and 10 specimens of normal oral mucosa from normal control subjects. In addition, we will study the correlation between the number of CD4+CD25+FOXP3+ Tregs and OLP subtypes or the disease activity of OLP.口腔扁平苔癬自體免疫疾病FOXP3+調節性T細胞γ-干擾素oral lichen planusautoimmune diseaseFOXP3+ Treg cellsinterferon-gamma