HPV18 E7抗原T细胞表位的鉴定及CD4^+T淋巴细胞的免疫反应  

Identification of specific T lymphocyte epitope on E7 antigen of human papillomavirus type 18 and CD4^+ T-cell response

作　　者：余剑琴[1] 徐云升[2] 欧荣英[1] 张乾[1] 郑飞云[1] 

机构地区：[1]温州医科大学附属第一医院妇产科,325000 [2]温州医科大学附属第一医院皮肤科,325000

出　　处：《浙江医学》2015年第10期823-826,共4页Zhejiang Medical Journal

基　　金：国家自然科学基金项目(30500537)

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(HPV18E780-94)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<O.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。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.

关 键 词：人乳头瘤病毒 E7抗原 CD4^+T淋巴细胞 TH表位 

分 类 号：R392[医药卫生—免疫学]