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目的:初步筛选湖北省五峰县土家族健康妇女HPV16型E7抗原HLA-A*02限制性CTL表位肽。方法:采集湖北省五峰县土家族HLA-A*02健康妇女外周全血并分离单个核细胞(peripheral blood mononuclear cell,PBMC),以HPV16型E7肽作为抗原,与人PBMC共培养9d后,采用酶联免疫斑点法(enzyme-linked immunosorbent spot,EILSPOT)检测不同多肽刺激PBMC产生特异性免疫反应的差异,使用SPSS 16.0软件进行统计学分析。结果:实验组均能诱导PBMC产生特异性免疫应答,其中E741-55(PAGQAEPDRAHYNIV)和E751-65(HYNIVTFCCKCDSTL)免疫反应显著。经过方差分析,组间差异有统计学意义,F=155.3,P=0.001;对组间进行LSD检验,与阴性对照组相比,实验组差异均有统计学意义,P=0.003;其中,P2和P4与阴性对照组比较差异有统计学意义,P值分别为0.021和0.016。结论:E741-55(PAGQAEPDRAHYNIV)和E751-65(HYNIVTFCCKCDSTL)抗原性强,有可能作为该地区HPV感染治疗性肽疫苗的候选表位。
OBJECTIVE: To screen HLA-A * 02 restricted CTL epitope peptide of HPV16 E7 antigen in healthy Tujia women in Wufeng County of Hubei Province. Methods: Peripheral blood mononuclear cells (HLA-A * 02) were collected from Tujia HLA-A * 02 healthy women in Wufeng County of Hubei Province. Peripheral blood mononuclear cells (PBMC) were isolated. HPV16 E7 peptide was used as antigen and co-cultured with human PBMC for 9 days. The difference of specific immune response induced by different peptides stimulated by different peptides was detected by enzyme-linked immunosorbent spot (EILSPOT), and the statistical analysis was carried out by using SPSS 16.0 software. Results: All the experimental groups could induce PBMC to produce specific immune response. E741-55 (PAGQAEPDRAHYNIV) and E751-65 (HYNIVTFCCKCDSTL) had significant immunoreactivity. After analysis of variance, the difference between groups was statistically significant, F = 155.3, P = 0.001; LSD test between groups, compared with the negative control group, the experimental group differences were statistically significant, P = 0.003; And P4 and the negative control group, the difference was statistically significant, P values were 0.021 and 0.016. CONCLUSION: E741-55 (PAGQAEPDRAHYNIV) and E751-65 (HYNIVTFCCKCDSTL) are highly antigenic and may serve as potential epitopes for therapeutic peptide vaccines for HPV infection in the region.