不同佐剂对人乳头瘤病毒16型L2E7E6蛋白免疫效果的影响  

Effects of Different Adjuvant on the Immune Responses to Recombi nant Protein L2E7E6 of Human Papillomavirus Type 16 in Mice

作　　者：任皎[1] 赵莉[1] 高孟[2] 姜云水[2] 郝明强[1] 谭文杰[1] 田厚文[1] 阮力[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京102206 [2]浙江省医学科学院病毒病研究所,杭州310013

出　　处：《生物技术通讯》2013年第3期337-341,共5页Letters in Biotechnology

基　　金：浙江省医学生物工程疫苗研究开发重点实验室开放课题

摘　　要：目的:研究CpG佐剂、弗氏佐剂、聚肌胞苷酸佐剂及左旋咪唑、西米替丁作为佐剂对人乳头瘤病毒16型L2E7E6融合蛋白在小鼠体内产生的免疫效果的影响。方法:以单独蛋白组、蛋白加各佐剂组分别肌肉注射免疫C57BL/6小鼠,检测不同佐剂诱发小鼠产生的体液免疫和细胞免疫应答水平,并观察其对小鼠肿瘤生长的抑制作用。结果:各免疫组均能检测到高滴度的抗L2、E7、E6蛋白IgG抗体(以IgG1为主),其中弗氏佐剂能显著提高E6蛋白的IgG和IgG1抗体水平和E7蛋白的IgG1抗体水平(P<0.05),CpG佐剂明显提高了E7蛋白的IgG2a抗体水平(P<0.01);而西米替丁佐剂则降低了E7抗原的IgG抗体水平(P<0.05);同时可以检测到CpG佐剂组能诱发小鼠产生针对E7、E6较强的细胞免疫反应,且能抑制70%的荷瘤小鼠肿瘤生长;此外弗氏佐剂与聚肌胞苷酸佐剂可产生较弱的针对E7肽的细胞免疫反应,能延缓荷瘤小鼠肿瘤形成时间,与单纯蛋白组相比差异显著(P<0.05)。结论:CpG佐剂、弗氏佐剂和聚肌胞苷酸佐剂都能提高人乳头瘤病毒16型L2E7E6融合蛋白的细胞免疫反应水平和抑制肿瘤生长能力,其中CpG佐剂效果较好,为促进该蛋白作为疫苗的研发提供了实验依据。Objective： To investigate the effects of CpG, freund adjuvant, poly I：C, levamisole and cimetidine on the immune responses to recombinant protein L2E7E6 of human papillomavirus type 16 in mice. Methods： C57BL/ 6 mice were immunized with recombinant protein L2E7E6 combined with or without CpG, freund adjuvant, poly I： C, levamisole or cimetidine through intramuscular. Then the humoral and cellular immune responses were detected by ELISA and ELISPOT respectively, and the effects of tumor growth regression were measured by using the TC-1 tumor bearing mice model. Results： All of groups mice immunized with the recombinant protein L2E7E6 combined with or without adjuvant could elicit high specific IgG antibodies titer against L2, E7 and E6 proteins （IgG1 antibodies were predominant）, among which those with freund adjuvant could secret higher IgG and IgG1 antibodies against E6 antigen and higher IgG1 antibody against E7 antigen respectively（P〈0.05）, and those with CpG could secret higher Thl-associated IgG2a antibodies against E7 antigen（P〈0.05）, however those with cimetidine could secret lower IgG antibody against E7 antigen（P〈O.05）. Furthermore, the mice vaccinated with the recombinant protein combined with CpG could elicit stronger T cell immune responses to peptides E7 and E6 respectively, and eliminate the established TC-1 tumors in 70% vaccinated mice. In addition, those mice vaccinated with freund adjuvant and poly I：C were able to elicit moderate cellular immune responses to peptide E7 and delayed tumor appearance in tumor-bearing mice. Conclusion： Accompanied with CpG, freund adjuvant or poly I：C, the recombinant protein could be enhanced on the cellular immune response and the effects of tumor growth regression in immunized mice, and it could have better effects with the CpG, which will provide the basis to develop the L2E7E6 protein as one promising candidate vaccine.

关 键 词：人乳头瘤病毒 佐剂 治疗性疫苗 

分 类 号：R392.1[医药卫生—免疫学]