Measurement of HIV Neutralization Using Quantitative Assays

在JC53-BL(TZM-bl)细胞中，通过荧光素酶报告基因分析，筛选出单个gp160克隆为环境假型病毒。对重组gp120疫苗的功能性包膜蛋白克隆进行了测序，并用可溶性CD4、单抗和重组gp120疫苗的感染者和非感染者的血清样本表征了它们的中和表型。来自不同地理位置的HIV-1分离株的环境克隆被挑选出来，这些克隆对中和并不异常敏感或抵抗，包括广泛的遗传、抗原和地理多样性。这些参考试剂将促进熟练程度测试和其他验证工作，旨在改善实验室间的测试性能，并可用于对疫苗引发的中和抗体进行标准化评估。 使用上述参考病毒试剂，BSL3核心病毒学实验室筛选了VRC研究人员产生的数百种动物血清。

Individual gp160 clones were screened for infectivity as Env-pseudotyped viruses in a luciferase reporter gene assay in JC53-BL (TZM-bl) cells. Functional env clones were sequenced and their neutralization phenotypes characterized by using soluble CD4, monoclonal antibodies and serum samples from infected individuals and non-infected recipients of a recombinant gp120 vaccine. Env clones from geographically diverse HIV-1 isolates were selected that were not unusually sensitive or resistant to neutralization and comprised a wide spectrum of genetic, antigenic and geographic diversity. These reference reagents will facilitate proficiency testing and other validation efforts aimed at improving assay performance across laboratories and can be used for standardized assessments of vaccine elicited neutralizing antibodies. Using the reference virus regents described above, the BSL3 core virology lab has screened hundreds of animal sera generated by investigators at the VRC.