人乳头状瘤病毒(HPV)16L1病毒样颗粒蛋白与HPV16L1基因联合免疫的体液免疫反应及其抗体的体外中和实验

Humoral immune response and in vitro neutralizing antibody assay on co-delivery of protein HPV16L1 virus-like particle with HPV16L1 gene

目的用含有编码人乳头状瘤病毒(HPV)16L1和HPV16L1病毒样颗粒(VLP)蛋白联合免疫小鼠,与用HPV16L1重组表达质粒比较,观察VLP蛋白免疫对免疫小鼠产生抗体的增强作用,以及抗体的体外中和作用,寻找研制HPV16感染的预防性疫苗的有效途径。方法将C57BL/6小鼠,随机分为4组:Ⅰ组:pcDNA-L1(100μl/鼠),Ⅱ组:pcDNA-L1(70μl/鼠)+HPV16L1VLP,Ⅲ组:pcDNA3.1(100μl/鼠)空质粒,Ⅳ组:PBS缓冲液。质粒免疫3次,间隔3周。ELISA法检测其血清抗体,红细胞凝集实验和HPV16病毒样颗粒结合抑制实验体外检测抗体的中和活性。结果pcDNA-L1+HPV16L1VLP较pcDNA-L1免疫组,3次免疫后的抗体水平均增高,尤其以第2次和第3次免疫后的抗体水平增加更显著。pcDNA-L1+HPV16L1VLP联合免疫组血清的抑制活性高于pcDNA-L1免疫组,且HeLa细胞结合抑制实验染色呈阴性。结论HPV16L1VLP联合免疫可以增加目的抗原的中和抗体产生,可能是HPV16有效预防性疫苗研制的更有希望的策略。

Objective To compare humoral immune response by co-inoculating mice with antigen HPV16L1 virus-like particle (VLP) and HPV16L1 recombinant plasmids and then observing the neutralizing antibody activity in vitro. Methods C57BL/6 mice were injected intramuscularly/subcutaneously with pcDNA-L1 plasmids plus HPV16L1 VLP. Serum IgG levels were detected by ELISA, antibody neutralizing protective activities were determined by hemagglutination inhibition and HPV16L1 VLP binding inhibition assay. Results Serum antibody titers and neutralizing antibody activities were increased in HPV16L1 plasmids plus HPV16L1 VLP proteins in co-immunized mice when compared with controls. Conclusion Co-inoculation of the HPV16L1 VLP protein can enhance production of neutralizing antibody activities against aimed antigen, which should be a more promising strategy for effective HPV16 prophylactic vaccine development.