摘要
目的 了解人乳头瘤病毒 (HPV) 6b型病毒样颗粒 (VLP)的免疫活性及其诱导的保护性抗体对HPV11型VLP和牛乳头瘤病毒 1型 (BPV1)VLP的交叉免疫反应。方法 将HPV6b、HPV11和BPV1晚期基因L1的开放读码框架 (ORFs)分别重组到杆状病毒中 ,该重组病毒在感染的昆虫细胞 (Sf 9细胞 )中表达 ,表达的L1蛋白可自行组装成HPV6b、HPV11和BPV1VLP。取 5 0 μg纯化的HPV6bVLP分别在第 0天和第 2 1天经肌肉免疫Balb/c鼠。第二次免疫后 1周及 3个月取血 ,检测血清抗HPV6bVLP、HPV11VLP和BPV1VLPIgG滴度 ;血凝集抑制试验检测HPV6bVLP免疫产生的抗血清是否能阻止HPV11VLP和BPV1VLP与鼠红细胞凝集。结果 免疫后 1周 ,应用ELISA方法检测抗HPV6bVLP、HPV11VLP和BPV 1VLP血清IgG滴度分别为 1∶6 4 0 0、1∶16 0 0和 1∶16 0 0。 3个月后 ,抗HPV6bVLP、HPV11VLP和BPV 1VLP血清IgG滴度分别为 1∶80 0、1∶4 0 0和 1∶10 0。血凝集抑制试验显示HPV6bVLP产生的抗血清能够阻止高度同源性的HPV6bVLP和HPV11VLP与鼠红细胞结合。结论 HPV6bVLP具有很强的免疫原性 ,免疫后产生的抗血清具有阻止HPV6bVLP和HPV11VLP与细胞结合的能力。HPV6b和 11型间确实存在交叉反应的中和表位 。
Objective To confirm human papillomavirus (HPV) 6b virus like particles (VLP) have strong immunogenicity and the protective antibody induced by HPV 6b VLP have cross reactive immunity against HPV11 VLP and bovine papillomavirus (BPV) 1 VLP. Method The late gene L1 for HPV6b, HPV 11 and L1/L2 for BPV 1 were molecularly cloned into recombinant baculovirus, respectively. The recombinant viruses were expressed in insect cells (Sf 9 cells). The expressed L1 proteins self assembled into virus like particles (VLP) for HPV6b, HPV 11 and BPV 1. VLP were purified from insect cell nuclei by CsCl centrifugation. The Balb/c mice were immunized on days 0 and 21 with 50 μg HPV6b VLP intramuscularly. Sera were collected after a further 7 days and 3 months. The titers of IgG against HPV 6b VLP, HPV 11 VLP and BPV1 VLP were detected. Hemagglutination inhibition assay was connducted to detected that whether antisera produced by HPV 6b VLP immunization could inhibit HPV11 VLP and BPV 1 VLP agglutinate mouse red blood cells. Result After 7 days of two immunizations, the titers of IgG against HPV6b VLP, HPV11 VLP and BPV1 VLP were 1∶6 400, 1∶1 600 and 1∶1 600 by ELISA, respectively. Three months later, the titers of IgG against HPV6b VLP, HPV11VLP and BPV1 VLP were 1∶800,1∶400 and 1∶100, respectively. Hemagglutination inhibition assay results showed that the antisera produced by HPV6b VLP inhibit HPV6b VLP and HPV11 VLP to mouse red blood cells binding. Conclusion HPV 6b VLP have potent immunogenicity. Antisera produced by HPV6b VLP could inhibit the binding of HPV6b VLP and HPV11 VLP and cells. Both HPV6b and HPV11 share neutralizing epitopes which are cross reactive and HPV6b VLP may be used in prophylactic and therapeutic vaccine for HPV6b and/or HPV 11 infections.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2002年第9期587-589,共3页
National Medical Journal of China
