粉尘螨3类变应原的T细胞表位预测及鉴定

目的 目的 预测并鉴定粉尘螨3类变应原 （Der f 3） 的T细胞表位。方法 方法 运用生物信息学分析软件预测Der f 3的T细胞抗原表位， 并人工合成所预测的T细胞表位肽。采用改良MTT法， 用预测表位肽刺激致敏鼠脾淋巴细胞进行脾淋巴细胞增殖试验； 酶联免疫吸附试验检测脾细胞培养上清液中白细胞介素?2 （IL?2）、 γ干扰素 （IFN?γ）、 IL?4及IL?5水平。结 结果 果 成功预测了Der f 3的5个T细胞表位肽， 通过脾淋巴细胞增殖试验， 其中3个表位肽序列可促进脾淋巴细胞增殖并刺激细胞因子IL?2和IFN?γ的分泌， 抑制细胞因子IL?4和IL?5的分泌。其序列分别为37GDCPYQISLQSSSHFCGG54、 98IY?QHENYDSMTIDNDVALIKLKTPMT123和164SELQRVDIDVVSREQCDQLYS184。结论 结论 初步鉴定了Der f 3变应原中3个T细胞表位序列， 为后续过敏性哮喘的诊断和特异性免疫治疗奠定基础。

Prediction and identification of T-cell epitopes in major group 3 allergen derived from Dermatophagoides farina

Objective To predict and identify T cell epitopes of major group 3 allergen derived from Dermatophagoides fari?na（Der f 3） . Methods The T cell epitopes of Der f 3 were analyzed through the sequence analysis by using the bioinformaticsonline tools. The five predicted peptides of T?cell epitopes were artificially synthesized. The spleen lymphocytes were co?culturedwith the five T cell epitopes by using the modified MTT method and the levels of IL?2，IFN?γ，IL?4 and IL?5 in the supernatantof the cultures were detected by ELISA. Results Five T cell epitopes of Der f 3 were predicted and three of which could pro?mote the proliferation of the mouse spleen lymphocytes. The secretions of IL?2 and IFN?γ were significantly induced and the se?cretions of IL?4 and IL?5 were significantly decreased by three of five prediction epitopes of Der f 3：37GDCPYQISLQSSSHFC?GG54，98IYQHENYDSMTIDNDVALIKLKTPMT123and 164SELQRVDIDVVSREQCDQLYS184. Conclusion Three T cell epitopesof Der f 3 have been initially identified，which lays the foundation of the diagnosis and treatment of asthma.