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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、98IYQHENYDSMTIDNDVALIKLKTPMT123和164SELQRVDIDVVSREQCDQLYS184。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。
Objective To predict and identify the T cell epitopes of der f 3 allergens (Der f 3). Methods The T cell epitopes of Der f 3 were predicted by bioinformatics analysis software and the predicted T cell epitope peptide was synthesized. The modified MTT method was used to test the splenic lymphocyte proliferation of sensitized murine splenic lymphocytes with the predicted epitope peptide. The levels of interleukin-2 (IL-2), γ in the culture supernatant of splenocytes were detected by enzyme-linked immunosorbent assay (IFN-γ), IL-4 and IL-5 levels. Results Five T-cell epitope peptides of Der f 3 were successfully predicted. Three epitope peptide sequences could promote the proliferation of spleen lymphocytes and stimulate the secretion of cytokines IL-2 and IFN-γ by spleen lymphocyte proliferation test. Inhibit the secretion of cytokines IL-4 and IL-5. The sequences were 37GDCPYQISLQSSSHFCGG54, 98IYQHENYDSMTIDNDVALIKLKTPMT123 and 164SELQRVDIDVVSREQCDQLYS184, respectively. Conclusion Preliminary identification of three T cell epitopes in the Der f 3 allergen provides a basis for the diagnosis and specific immunotherapy of subsequent allergic asthma.