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目的制备含人乳头瘤病毒(humanpapillomavirus,HPV)58型E7基因的重组痘苗病毒疫苗,并利用动物肿瘤模型观察疫苗接种效果。方法利用PCR方法扩增HPV58E7DNA片段,在消除其转化活性的基础上,构建表达HPV58E7的重组痘苗病毒疫苗并免疫小鼠,观察该疫苗对E7阳性肿瘤细胞生长的抑制作用,体外诱导、测定细胞毒性T淋巴细胞(cytotoxicTlymphocytes,CTL)活性。结果经定点突变的HPV58E7基因转化活性显著降低,用突变的E7基因构建的重组痘苗病毒免疫小鼠后,能抑制E7阳性肿瘤细胞在小鼠体内的生长并延长小鼠的生存期;免疫小鼠脾淋巴细胞体外可诱导产生针对E7阳性肿瘤的CTL细胞。结论在消除转化活性的基础上,HPV58E7可用于治疗与HPV58感染有关的肿瘤。
Objective To prepare recombinant vaccinia virus vaccine containing HPV type E7 gene of human papillomavirus (HPV), and to observe the effect of vaccination by animal tumor model. Methods The HPV58E7 DNA fragment was amplified by PCR and the recombinant vaccinia virus vaccine expressing HPV58E7 was constructed and immunized to eliminate the transformation activity. The inhibitory effect of the vaccine on the growth of E7 positive tumor cells was observed, and the cytotoxicity T lymphocyte (CTL) activity. Results The site-directed mutagenesis of HPV58E7 gene was significantly reduced. Immunization of mice with the recombinant vaccinia virus with the mutant E7 gene inhibited the growth of E7-positive tumor cells in mice and prolonged the survival of mice. Immunized mice Spleen lymphocytes can induce the production of CTL cells against E7 positive tumors in vitro. Conclusion Based on the elimination of transformation activity, HPV58E7 can be used to treat tumors associated with HPV58 infection.