粉尘螨3类变应原的T细胞表位预测及鉴定

发布时间：2018-11-28 17:25

【摘要】：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。
[Abstract]:Objective to predict and identify T cell epitopes of allergen (Der f 3 of Dermatophagoides farinae. Methods the T cell epitopes of Der f 3 were predicted by bioinformatics analysis software, and the predicted T cell epitope peptides were synthesized. The spleen lymphocyte proliferation test was carried out by using the modified MTT method to stimulate the spleen lymphocytes of the sensitized mice with predicted epitope peptide. The levels of interleukin-2 (IL-2), interferon 纬 (IFN- 纬), IL-4 and IL-5 in the supernatant of splenocyte culture were detected by enzyme-linked immunosorbent assay (Elisa). Results five T cell epitope peptides of Der f 3 were successfully predicted. By splenocyte proliferation test, three epitope peptide sequences could promote the proliferation of spleen lymphocytes and stimulate the secretion of cytokines IL-2 and IFN- 纬. Inhibit the secretion of cytokines IL-4 and IL-5. The sequences were 37GDCPYQISLQSSHFG5FG5FN 98IYQHENYDSMTIDNDVALIKLKTPMT123 and 164SELQQRVDIDVSREQLVSREQLYS184. Conclusion three T cell epitope sequences of Der f 3 allergen were preliminarily identified, which laid a foundation for the diagnosis and specific immunotherapy of allergic asthma.
【作者单位】：皖南医学院医学寄生虫学教研室;皖南医学院生理学教研室;
【基金】：国家自然科学基金(81270091、30872367、81172790)
【分类号】：R392

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