Measurement of HIV Neutralization Using Quantitative Assays

在JC 53-BL（TZM-bl）细胞中的荧光素酶报告基因测定中，筛选单个gp 160克隆作为Env假型病毒的感染性。 对功能性env克隆进行测序，并通过使用可溶性CD 4、单克隆抗体和来自重组gp 120疫苗的感染个体和非感染接受者的血清样品来表征其中和表型。 选择来自地理上不同的HIV-1分离株的Env克隆，这些克隆对中和反应不具有异常敏感性或抗性，并且包含广泛的遗传、抗原和地理多样性。 这些参比试剂将促进能力验证和其他验证工作，旨在提高各实验室的检测性能，并可用于疫苗引起的中和抗体的标准化评估。 使用上述参考病毒试剂，BSL 3核心病毒学实验室筛选了VRC研究人员产生的数百份动物血清。

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.