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目的:探讨人乳头瘤病毒16型L1-E7重组腺病毒载体疫苗的免疫效应。方法:①将rAd5HPV16L1-E7载体疫苗在293细胞中扩增后,以肌肉注射、腹腔注射、滴鼻和灌胃途径免疫小鼠,采用ELISA法测其血清IgG抗体水平;②制备脾细胞悬液,加入到96孔无菌培养板中,并分别加入rAd5HPV16L1-E7特异性蛋白和RPMI1640+10%胎牛血清,加入3H-TdR1μCi/孔,做T-细胞增殖实验;③取经过培养56h的培养液进行IFN-γ的测定。结果:不同途径免疫的BALB/c小鼠血清IgG抗体水平均高于对照组(P<0.05),但以肌注组产生抗体量最多,IFN-γ和T-细胞增殖亦高于对照组(P<0.05)。结论:rAd5HPV16L1-E7重组活病毒疫苗既能刺激T-细胞产生细胞介导的免疫应答,又能刺激B-细胞产生抗体介导的免疫应答,说明其具有很好的免疫原性。
Objective: To investigate the immune effect of human papillomavirus type 16 L1-E7 recombinant adenovirus vector vaccine. Methods: (1) After the rAd5HPV16L1-E7 vector vaccine was amplified in 293 cells, the mice were immunized by intramuscular, intraperitoneal, intranasal and intragastric administration, and the serum IgG antibody levels were measured by ELISA. ② The spleen cell suspension , Added into 96-well sterile culture plate and added with rAd5HPV16L1-E7 specific protein and RPMI1640 + 10% fetal bovine serum, added 3H-TdR1μCi / well, do T-cell proliferation experiments; ③ culture 56h Fluid for the determination of IFN-γ. Results: The serum levels of IgG in BALB / c mice immunized with different routes of immunization were higher than those in control group (P <0.05), but the amount of antibody produced by intramuscular injection was the highest, and the proliferation of IFN-γ and T-cells was also higher than that of the control group P <0.05). CONCLUSION: The rAd5HPV16L1-E7 recombinant live virus vaccine can stimulate T-cells to produce cell-mediated immune responses and stimulate B-cells to produce antibody-mediated immune responses, indicating that they have good immunogenicity.