HPV16型E6蛋白T细胞抗原表位的变异致机体免疫应答差异及宫颈癌发病的机制研究

新疆地区宫颈癌发病率是内地人群的4~5倍，课题组前期研究发现新疆人群HPV16型感染率高（>85%）且以欧洲亚型(>95%)为主，其致癌基因E6存在L83V的高频突变（58.1%），并首次报道了D64E（8.7%）突变，这两个突变共造成15个T细胞抗原表位变异，其是否造成了免疫逃逸并致宫颈癌尚未有报道。本项目拟收集HPV16型感染的宫颈癌组、持续感染组和一过性感染组，Taqman探针法验证HPV16型的突变分布，分析L83V、D64E突变与HPV16型持续感染及致宫颈癌进展的相对危险；合成L83V、D64E对应的15个TCE野生型、突变型的抗原肽，Elispot和FCM实验检测对应患者γ干扰素等细胞因子释放水平，验证T细胞抗原表位变异导致个体迟发性超敏反应的差异，本项目有望发现HPV16型基因突变致免疫逃逸和宫颈癌进展的机制，以期揭示新疆人群宫颈癌高发的原因，为宫颈癌疫苗应用提供理论依据。

The incidence of cervical cancer in Xinjiang population is 4~5 times that of the mainland population. We found that the HPV16 infection rate is high (>85%) in Xinjiang population and is mainly European subtypes (>95%). Its oncogene E6 has a high-frequency mutation of L83V(58.1%), and the mutation of D64E (8.7%) was reported for the first time. These two mutations caused a total of 15 T cell epitope (TCE) mutations, whether it caused immune escape and cervical cancer has not been reported. This project sets up HPV16-infected cervical cancer group, persistent infection group and transient infection group, 60 patients in each group, Taqman probe method was used to verify the mutations distribution of HPV L83V and D64E, and to analyze the relative risk of mutations and persistent HPV16 infection and cervical cancer; Synthesize 15 TCE wild-type peptides and mutant antigen peptides of L83V and D64E, Elispot and FCM experiments detect the release levels of cytokines such as interferon in the patients; analyze the differences in individual delayed type hypersensitivity caused by TCE variation; Clarify the mechanism of HPV16 TCE mutation-induced immune escape. The implementation of the project is expected to reveal the reasons for the high incidence of cervical cancer in Xinjiang population, and provide a theoretical basis for the application of cervical cancer vaccine.