一种直接评价HPV16L1抗体活性的新方法

目的 :以pcDNAL1质粒免疫啮齿类动物 (C5 7BL/ 6 )为模型 ,观察HPV16L1VLP细胞结合抑制实验是否可以用于检测免疫抗体的中和保护作用。方法 :实验组包括Ⅰ组pcDNAL1、Ⅱ组HPV16L1VLP ;Ⅲ组pcDNA3.1。每组动物均为 6只C5 7BL/ 6鼠。每组动物均肌肉注射免疫 3次 ,间隔 3w。末次免疫后 14d眼球后取血并拉颈处死动物 ,进行HPV16L1VLP结合抑制试验 :免疫血清中和HPV16L1VLP ;制备Hela ,EJ和RLC310细胞爬片 ,CS12 13细胞涂片 ;细胞免疫组化染色。结果 :实验组血清中和后Hela细胞呈染色阴性 ,而对照组、不共育组则细胞呈棕黄色。结论 :这说明实验组血清具有抑制VLP与Hela细胞粘附的活性效应。这一方法可能比HAI更能直接反映中和抗体的活性和保护作用。

The method of directly assessing the immune-protective activity of HPV16L1 antibody

Objective:To observe whether the HPV16L1VLP Hela-cell binding-inhibition experiment can assess the immune-protective activities of HPV16L1 antibody in HPV16 prophylactic vaccine or other fields.Methods:C57BL/6 mice were divided into three groups,including pcDNAL1 plasmid,HPV16L1VLP protein,pcDNA3.1 plasmid.Each group of mice was immunized intramuscularly or subcutaneously with certain above antigens,accompanying three times and interval 3 weeks.After 14 days of 3 rd immunization,take blood to make serum for immunochemistry.Results:Hela cell neutralized by the serum IgG of experimental mice can be stained negatively,while the cells of control and non-inoculation groups can be stained positively (brown-yellow color).Conclusion:HPV16LIVLP Hela-cell binding-inhibition assay may be the better way to reflect the neutralizing-protective activities of HPV16L1 antibody in many fields including HPV16 prophylactic vaccine.