中文摘要：

目的初步鉴定人乳头瘤病毒（HPV）18型E7抗原的辅助T淋巴细胞（Th）表位及CD4^＋T淋巴细胞的免疫反应。方法以免疫磁珠进一步分离HLA—DRB1^＊0301阳性健康人外周血单核细胞（PBMC）中的CD4^＋T淋巴细胞，流式细胞仪鉴定细胞纯度，用之前实验中已获得的3个HPV18 E7抗原HLA—DRB1^＊0301限制性Th表位刺激外周血CD4^＋T淋巴细胞，用5-溴脱氧尿嘧啶核苷（Brdu）检测CD4^＋T淋巴细胞增殖活性。ELISA方法分析表位刺激CD4^＋T淋巴细胞分泌的细胞因子。结果多肽P1（HPV18 E780—94）在体外刺激CD4^＋T淋巴细胞后能够有效诱导CD4^＋T淋巴细胞增殖，P1组与阴性对照组比较差异有统计学意义（P〈0．01），多肽P1（HPV18 E780—94）刺激CD4^＋T淋巴细胞分泌IFN—γ，P1组与阴性对照组比较差异有统计学意义（P〈0．01）。结论P1（HPV18 E780—94）可能为HPV18 E7抗原HLA—DRB1^＊0301限制性Th表位。

英文摘要：

Objective To identify specific T lymphocyte epitope on E7 antigen of human papillomavirus type 18 and CD4^＋ T-cell responses. Methods Three epitope candidates of human papillomavirus 18 E7 antigen （P1, P2 and P3） were obtained previously. Peripheral blood mononuclear cell （PB MC） from HLA- DRB1^＊0301-positive healthy donors were prepared and CD4^＋ T cells were separated by Dynal Immunomagnetic beads isolation. The purity was determined by flow cytometry. The predicted peptides were scored as immunogenic ability to stimulate CD4^＋ T cells. Proliferative activity was analyzed with Brdu cell proliferation assay. The cytokine production of CD4^＋ T cells induced by the peptides was detected by ELISA. Results Peptide P1 （HPV18 E780-94） stimulated proliferation of CD4^＋T cells and the OD value was significantly higher than that of negative control group（P〈0.01）. Peptide P1 （HPV18 E780-94） also stimulated CD4^＋T cells to secrete IFN-γ and IFN-γ, value was significantly higher than that of negative control group（P〈0.01）. Conclusion Peptide P1（HPV18 E780-94） can be the HLA-DRB1^＊0301-restricted Th epitope of human papillomavirus 18 E7 antigen, which provides the basis to evaluate cellular immune response elicited by HPV18 E7 protein based vaccine.