寻常型天疱疮自身抗原Dsg3片段的重组表达和特异性细胞反应

目的 :探讨寻常型天疱疮自身抗原Dsg3在特异性T细胞反应中的作用 ,为自身免疫性疾病机制的研究提供依据。方法 :根据Genbank中的Dsg3序列分析 ,采用RT PCR法克隆自身抗原Dsg3E1,E2 ,E3,E4,E5多肽片段的cDNA ,定向插入表达载体PGEX 2T ,导入大肠杆菌JM10 9中表达重组融合蛋白并经GST层析柱纯化 ;进一步与PV患者及疾病对照组、正常对照组T细胞混合培养 ,观察T细胞增殖反应。结果 :Dsg3E1,E2和E4,E5可刺激PV患者T细胞反应 ,而不与疾病对照组、正常对照组反应。结论 :Dsg3E1,E2和E4,E5中包含T B细胞作用相关的抗原表位 ,在PV发病中起重要作用。

Expression and specific cell response of autoantigen Dsg3 in pemphigus vulgaris

Objective:In view of analysis of role of autoantigen Dsg3 in specific T cell response to understand the molecular mechanism of autoimmune diseases.Methods:Based on the sequence analysis of autoantigen Dsg3,the five fractions cDNAs of Dsg3 were cloned and Dsg3-GST fusion proteins induced by IPTG in E.coli.JM109 were purified,then mixed-cultured with T cell from PV patients.The T cell proliferation response were analyzed by MTT.Results:Dsg3 E1,E2 and E4,E5 stimulated T cell from PV patients,not controls.Conclusion:Dsg3 E1,E2 and E4,E5 contained the epitopes relevant to T-B cell interaction,which play an important role in the pathogenesis of pemphigus vulgaris.