Objective To analyze Epstein-Barr virus(EBV) protein epitope polypeptide sequence and its human leukocyte antigen(HLA)genotypes with high frequency in Chinese.Methods A total of 23 polypeptides of EBV protein epitope were designed by NetMHC software. According to HLA phenotypes,HLA-A*2402,HLA-A*1101 and HLA-B*4001,polypeptides were classified into 3 groups. Peripheral blood mononuclear cells(PBMC) collected from 160 healthy subjects were stimulated with 3 groups of polypeptides. Interferon-gamma(IFN-γ) levels were determined by IFN-γ enzyme-linked immunosorbent assay(ELISA). CD8,interleukin 17(IL-17), CD40L,tumor necrosis factor alpha(TNF-α),CD45RA and Foxp3 expression levels were determined byflow cytometry.Results In 160 peripheral blood samples,61 cases were positive,of which 24 cases with HLA-A*2402 positive(15%),34 cases were HLA-A*1101 positive(21%),31 cases were HLA-B*4001 positive(19%). The expression levels of CD8,IL-17,CD40L and TNF-α in peripheral blood in positive reaction group were higher than those in negative reaction group(P<0.05),but the expression levels of CD45RA and Foxp3 had no statistical significance(P>0.05).Conclusions In Chinese,HLA-A*1101 genotype is dominant in EBV protein epitope polypeptide sequence.%目的 分析中国人群高频率的人类白细胞抗原(HLA)基因型及其所递呈EB病毒(EBV)蛋白表位序列.方法 通过NetMHC软件分析预测并合成HLA能够递呈EBV蛋白表位的23种多肽,按照HLA表型HLA-A*2402、HLA-A*1101和HLA-B*4001分为3组.收集160名健康志愿者的外周血单个核细胞(PBMC),用3组多肽刺激PBMC,血液分析仪检测增殖,采用γ干扰素(IFN-γ)酶联免疫吸附试验(ELISA)试剂盒检测IFN-γ水平,并用流式细胞术检测CD8、白细胞介素17(IL-17)、CD40L、肿瘤坏死因子α(TNF-α)、CD45RA、转录因子(Foxp3)的表达水平.结果 在160例外周血样本中,对3组多肽刺激反应阳性数为61例,其中HLA-A*2402型阳性数为24例,阳性率为15%;HLA-A*1101型阳性数为34例,阳性率为21%;HLA-B*4001型阳性数为31例,阳性率为19%.阳性反应组外周血中CD8、IL-17、CD40L、TNF-α的表达水平显著高于阴性反应组,差异有统计学意义(P<0.05),而CD45RA、Foxp3的表达水平差异无统计学意义(P>0.05).结论 中国人群HLA-A*1101基因型在递呈EBV表位序列中占优势.
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