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目的观察口服人乳头瘤病毒(HPV)18型E7联合次级淋巴组织趋化因子(SLC)基因疫苗的抗肿瘤作用。方法Western blotting方法鉴定稳定表达E7的B16F1细胞株。小鼠腋下接种稳定表达E7的小鼠黑色素瘤B16F1细胞,荷瘤小鼠随机分为5组(n=12),灌胃途径给予各种伤寒沙门菌液免疫小鼠:生理盐水组(生理盐水0.2mL)、空载体组(携带空载体的菌液0.2mL)、E7组(携带E7的菌液0.2mL)、SLC组(携带SLC的菌液0.2mL)和E7联合SLC组(携带E7或SLC的菌液各0.1mL)。检测各组小鼠免疫后的瘤重及生存率、血清及阴道分泌物E7特异性抗体产生情况、脾细胞增殖指数和杀伤活性,并观察肿瘤组织局部淋巴细胞浸润情况。结果稳定表达E7的B16F1细胞株成功构建。E7联合SLC组小鼠瘤重显著降低,生存期显著延长(P<0.05),诱导产生血清E7特异性IgG抗体水平显著增加(P<0.01),淋巴细胞增殖指数和细胞毒T淋巴细胞杀伤率显著高于其他组(P<0.05或P<0.01)。SLC组及E7联合SLC组肿瘤组织浸润的淋巴细胞数均明显增多。结论口服HPV18型E7联合SLC基因疫苗可发挥抗肿瘤作用,其机制可能为增强体液和细胞免疫应答。
Objective To observe the antitumor effect of oral HPV 18 E7 combined with lymphoid tissue chemokine (SLC) gene vaccine. Methods B16F1 cells stably expressing E7 were identified by Western blotting. Mice were inoculated with B7F1 cells stably expressing E7 mouse melanoma. The tumor-bearing mice were randomly divided into 5 groups (n = 12). All mice were immunized with Salmonella typhimurium by intragastric administration. Saline (0.2 mL), empty vector group (empty vector carrying 0.2 mL), E7 group (0.2 mL of E7-carrying bacterial solution), SLC group (0.2 mL of SLC-bearing bacterial solution) and E7 combined with SLC group Or 0.1 mL each of SLC bacterial liquid). Tumor weight and survival rate, serum and vaginal secretions E7-specific antibody production, spleen cell proliferation index and cytotoxic activity of the mice in each group were detected, and the local lymphocyte infiltration was observed. Results The B16F1 cell line stably expressing E7 was successfully constructed. The tumor weight of E7 combined with SLC group was significantly decreased (P <0.05), and the level of serum E7-specific IgG antibody was significantly increased (P <0.01). The lymphocyte proliferation index and cytotoxic T lymphocyte killing rate Significantly higher than other groups (P <0.05 or P <0.01). SLC group and E7 combined with SLC group tumor infiltration of lymphocytes were significantly increased. Conclusion Oral HPV18 E7 combined with SLC gene vaccine can play an antitumor effect, the mechanism may be to enhance humoral and cellular immune response.