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Abnormal DNA Methylation in CD4+T Cells from Patients with Primary Sj(?)gren’s Syndrome(pSS)

Author: YinHeng
Tutor: LuQianJin
School: Central South University
Course: Dermatology and Venereology
Keywords: CD70 T cells DNA methylation Primary Sjogren's syndrome
CLC: R589
Type: Master's thesis
Year: 2010
Downloads: 99
Quote: 1
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Abstract


Objective: Primary Sjogren's syndrome (primary sjogren's syndrome, pSS) is a lymphocytic infiltration, resulting in a large number of autoantibodies cause damage to the lacrimal and salivary glands characterized by autoimmune disease, its pathogenesis is not clear. In recent years, a growing number of studies have shown that epigenetic mechanisms, especially the T-cell DNA hypomethylation play a very important role in the pathogenesis of T-cell gene regulation and autoimmune disease. Such as systemic lupus erythematosus (systemic lupus erythematosus, SLE) showed overall genomic hypomethylation and gene-specific hypomethylation, the of CD11a (ITGAL,), of CD70 (TNFSF7), of CD40L (TNFSF5) and Perforin (PRF1) . T cells have over-expression of these genes autoreactive anti macrophage or help B cells to produce large amounts of autoantibodies play a very important role in the occurrence of SLE, the development. The main subject through research with primary Sjogren's syndrome CD4 T cells in the peripheral blood of patients with immune-related methylation sensitive gene expression levels and their methylation status of regulatory sequences, which explore pSS epigenetic changes, in order to reveals pSS pathogenesis open new avenues. Methods: 17 cases of pSS patients and 14 normal density gradient centrifugation of peripheral blood mononuclear cells by flow cytometry detected in PBMC CD11a, CD70 and CD40L protein expression levels in CD4 T cells; bead sorting CD4 T cells, quantitative real-time polymerase chain reaction (real-time RT-PCR) to detect mRNA expression of CD70 and Perforin; Western blot detection Perforin protein expression levels; sodium bisulfite sequencing to detect the expression of different gene regulatory sequences methyl of state. Results: Compared with the normal control group, pSS patients peripheral blood CD4 T cells, CD70 mRNA expression increased significantly (P = 0.001), protein expression levels also increased significantly (P = 0.003); compared with the normal control group, pSS patients peripheral blood CD4 T cells the Perforin of mRNA expression levels increased significantly (P = 0.018), while no significant difference in the level of protein expression. Compared with the normal control group of pSS patients with peripheral blood CD4 T-cell CD11a and CD40L expression levels had no significant difference. pSS patients with peripheral blood CD4 T cells, CD70 gene promoter methylation level significantly lower than the normal control group (P = 0.007). CD70 gene promoter DNA methylation level and CD70 mRNA expression levels showed a significant negative correlation (r = -0.698, P = 0.008). Conclusion: The CD70 gene promoter DNA hypomethylation may be involved in the regulation of primary Sjogren's syndrome patients with peripheral blood CD4 T cells, CD70 overexpression, suggesting that DNA methylation abnormalities may play in the pathogenesis of primary Sjogren's syndrome role.

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