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目的:构建共表达人乳头瘤病毒16型(HPV16)L1、L2、E67蛋白的非复制型重组痘苗病毒人用疫苗株。方法:以痘苗病毒为载体、利用同源重组技术筛选共表达HPV16L1、L2、E67蛋白的重组痘苗病毒并对其进行鉴定。结果:该病毒在CEF细胞上连续传至第15代,经斑点杂交结果表明重组病毒基因组中有L1、L2、E6、E7基因插入;经Westernblot检测,重组病毒能稳定表达HPV16L1、L2、E67蛋白。结论:非复制型重组痘苗病毒NTVJE67CKL1L2可作为预防和治疗HPV16相关肿瘤及其癌前病变的候选疫苗。
Objective: To construct a non-replicating recombinant vaccinia virus human vaccine strain coexpressing human papillomavirus type 16 (HPV16) L1, L2 and E67 proteins. Methods: The recombinant vaccinia virus co-expressing HPV16 L1, L2, and E67 proteins were screened by homologous recombination using vaccinia virus as a vector and identified. Results: The virus was continuously transmitted to passage 15 on CEF cells. The results of dot blot hybridization showed that L1, L2, E6 and E7 genes were inserted into the recombinant virus genome. The recombinant viruses could stably express HPV16 L1, L2 and E67 proteins by Western blot . Conclusion: The non-replicating recombinant vaccinia virus NTVJE67CKL1L2 can be used as a candidate vaccine for the prevention and treatment of HPV16-related tumors and precancerous lesions.