【摘要】 目的　研究重组人乳头瘤病毒 6型 (humanpapillomavirustype 6 ,HPV 6 )病毒样颗粒(virus likeparticle ,VLP)的免疫原性。方法　重组杆状病毒在昆虫细胞中表达制备的HPV 6L1VLP(L1 VLP)和HPV 6L1+L2VLP(L1+2 VLP)经鉴定后 ,用于免疫BALB/c小鼠 ,对诱导的体液免疫和细胞免疫反应进行了检测。结果　电镜观察显示L1 VLP和L1+2 VLP二者形态上无明显差异 ,为圆形颗粒 ,直径约 5 0nm ,SDS PAGE和Westernblot分析表明 ,L1+2 VLP中L1和L2蛋白摩尔比例为 4∶1。用ELISA法测定免疫小鼠血清抗体滴度 ,加佐剂L1 VLP免疫组和加佐剂L1+2 VLP免疫组血清针对HPV 6L1VLP的滴度在 1∶10 0 0 0以上 ,高于未加佐剂组免疫血清滴度 (1∶2 0 0 0 ) ,L1+2 VLP免疫诱导出了特异于L2抗原的抗体。血清抗体主要识别HPV 6构象依赖性抗原表位 ,与HPV 11抗原显示出一定的交叉反应 ,而与HPV 16无明显交叉反应。免疫小鼠脾淋巴细胞体外经HPV 6L1VLP再激活后出现了特异性增殖反应 ,3H TdR掺入值与未免疫组之间差异有显著性 (P <0 .0 1) ,L1 VLP和L1+2 VLP两组间差异无显著性 (P >0 .0 5 ) ,L1 VLP和L1+2 VLP免疫组刺激指数 (SI)分别为 6 .4和 6 .2 ,阴性对照组SI为 1.1。HPV 6L1VLP再刺激特异地诱导免疫组脾淋巴细胞IL 2和IL 10分?更多还原

【Abstract】 Objective To investigate the immunogenicity of recombinant human papillomavirus type 6(HPV-6) virus-like particles(VLPs). Methods HPV-6 L1 VLP(L1-VLP) and L1+L2 VLP(L1+2-VLP) were prepared from Sf9 cells that were infected with L1 recombinant baculovirus or co-infected with additional L2 recombinant baculovirus, and were characterized by SDS-PAGE, Western blot and electron microscopy. The L1-VLP and L1+2-VLP were used to immunize BALB/c mice, and the humoral and cellular immune responses were detected. Results L1-VLP and L1+2-VLP were both spherical particles with a diameter of approximately 50nm. L1+2 VLP comprised L1 and L2 in the molar ratio of 4∶1. The titers of serum antibodies in groups vaccinated with adjuvant L1-VLP and L1+2-VLP were above 1∶10?000 as detected by ELISA, which was higher than that in groups without adjuvant (1∶2?000), L1+2-VLP induced antibodies specific to L2 antigen. The serum antibodies mainly recognized confirmation-dependant HPV-6 epitopes, showing cross-reactivity to HPV-11 but not to HPV-16. Specific splenic lymphocytes proliferation after in vitro re-stimulating with HPV-6 L1 VLP was observed in 3H-TdR incorporation test for immunized mice (P<0.01), but not significant difference between L1-VLP and L1+2-VLP immunized group (P>0.05). The stimulation indexes(SI) were 6.4, 6.2 and 1.1 for L1-VLP, L1+2-VLP and control group. In vitro re-stimulation also specifically induced significant increase of IL-2 and IL-10 secretion in the splenic lymphocytes from VLP-vaccinated group (P<0.01). However, no significant increase was found for IFN-γ. Conclusion The prepared recombinant HPV-6 VLPs were highly immunogenic inducing both cellular and humoral immunity in mice, and can be used to explore preventative vaccine against HPV. [更多还原