【作者】 施桥发；

【导师】 李明远；

【作者基本信息】 四川大学 ， 病原生物学， 2006， 博士

【摘要】 目的 HPV16是引起子宫颈癌等疾病的重要原因。在子宫颈癌患者中，90％以上的患者存在HPV感染，其中60％以上为HPV16感染。在HPV16感染的子宫颈癌细胞中，HPV16 E7蛋白存在持续性表达，细胞内HPV16E7蛋白与肿瘤抑制蛋白Rb蛋白结合，使原本结合于Rb蛋白的真核细胞转录起始因子E2F释放，引起细胞内多种蛋白的异常表达，使细胞突破生长检查点由G1期向S期转变，是引起起细胞恶性转化的重要原因，因此被认为是HPV16相关肿瘤特异性抗原，也被广泛地认为是HPV16相关肿瘤免疫治疗的理想靶点之一。 研究表明HPV16感染相关的临床诊断主要依靠以PCR技术为主的分子生物学方法，该方法具有设备条件要求高、监测结果假阳性率高、标本采集不方便、不易操作和推广等缺点，而免疫学诊断方法具有易于操作、标本采集本方便、检测敏感性高、特异性好的特点。另一方面研究表明，HPV16感染患者体内存在微量HPV16 E7抗体，细胞内HPV16E7蛋白存在持续性表达，它们可以作为有效的免疫学检测靶。 因此，本研究一方面拟探索HPV16早期蛋白E7的免疫学特性，为以HPV16 E7蛋白作为靶点的免疫学治疗性疫苗研究提供线索，为HPV16相关恶性肿瘤的免疫辅助治疗提供依据；另一方面，以原核表达的HPV16 E7-TRX融合蛋白为抗原，制备抗HPV16 E7单克隆抗体，为HPV16感染相关疾病的免疫学诊断提供线索。更多还原

【Abstract】 ObjectivesHuman papillomavirus type 16(HPV16) is the most important cause for cervical cancer. Over 90% of cervical cancer patients have ever infected with HPV, more than 60% of which were texted as HPV16. After the genome of HPV16 integrated into celluar chromosome, HPV16 E7 protein can be persistently expressed with high level in it. HPV16 E7 protein can bind effectively with Rb, which is one of the most important tumor suppressor proteins for cell. In normal condition, Rb is bound with E2F, which is a major transcription initiation factor for eukaryocytes. Once at the time Rb is bound with HPV E7, not only will Rb protein be degradated quickly, but also E2F will be released greatly, it will finally enhance expression of many proteins which are not expressed at normal. All of these can bring the cell checkpoint from G1 to S phase, which resulting in the cell being a condition named malignant transformation. So it is a popular view looking HPV16 E7 as a tumor specific antigen(TSA) for HPV16 related cervical cancer and it is also confirmed as an ideal target for the immunotherapy on HPV16 related cancer.Up to date, PCR is a main weapon used to find HPV infection, including HPV16, which is most important in clinical. But as we know, PCR technique is greatly dependented on the equipment and it must be performed under a relatively seperated and clean environment. Otherwise it will give us many false positive information. On更多还原