a Division of HIV/AIDS and Sex-transmitted Virus Vaccines , National Institutes for Food and Drug Control (NIFDC) , Beijing , China.
b Beijing You'an Hospital, Capital Medical University , Beijing , China.
Hum Vaccin Immunother. 2018 Jan 2;14(1):199-208. doi: 10.1080/21645515.2017.1373922. Epub 2017 Oct 18.
Sensitive assays for HIV-1 neutralizing antibody detection are urgently needed for vaccine immunogen optimization and identification of protective immune response levels. In this study, we developed an easy-to-use HIV-1 pseudovirus neutralization assay based on a human CD4 lymphoblastoid cell line A3R5 by employing a high efficient pseudovirus production system. Optimal conditions for cell counts, infection time, virus dose and concentration of DEAE-dextran were tested and identified. For T-cell line-adapted tier 1 virus strains, significantly higher inhibitory efficiency was observed for both monoclonal neutralizing antibody (4 fold) and plasma (2 fold) samples in A3R5 than those in TZM-bl assay. For circulating tier 2 strains, the A3R5 pseudovirus assay showed even much higher sensitivity for both neutralizing antibody (10 fold) and plasma (9 fold) samples. When sequential neutralizing antibody seroconverting samples were tested in both A3R5 and TZM-bl assays, the seroconverting points could be detected earlier for tier 1 (15.7 weeks) and tier 2 (68.3 weeks) strains in A3R5 assay respectively. The high sensitive pseudovirus assay using more physiological target cells could serve as an alternative to the TZM-bl assay for evaluation of vaccine-induced neutralizing antibodies and identification of the correlates of protection.
迫切需要灵敏的 HIV-1 中和抗体检测分析方法，用于疫苗免疫原优化和鉴定保护性免疫反应水平。在这项研究中，我们开发了一种基于人 CD4 淋巴母细胞系 A3R5 的易于使用的 HIV-1 假病毒中和测定法，该方法采用了高效的假病毒生产系统。对细胞计数、感染时间、病毒剂量和 DEAE-葡聚糖浓度的最佳条件进行了测试和鉴定。对于 T 细胞系适应的 1 级病毒株，在 A3R5 中观察到单克隆中和抗体（4 倍）和血浆（2 倍）样本的抑制效率明显高于 TZM-bl 测定法。对于循环的 2 级株，A3R5 假病毒测定法对中和抗体（10 倍）和血浆（9 倍）样本的敏感性更高。当在 A3R5 和 TZM-bl 测定法中检测连续的中和抗体血清转换样本时，A3R5 测定法可分别更早地检测到 1 级（15.7 周）和 2 级（68.3 周）株的血清转换点。使用更具生理意义的靶细胞的高灵敏度假病毒测定法可替代 TZM-bl 测定法，用于评估疫苗诱导的中和抗体，并鉴定保护性相关因素。
