Measurement of HIV Neutralization Using Quantitative Assays

在JC53-BL （TZM-bl）细胞中进行荧光素酶报告基因检测，筛选gp160克隆个体作为env型病毒的感染性。对功能环境克隆进行了测序，并利用可溶性CD4、单克隆抗体和重组gp120疫苗感染者和非感染者的血清样本对其中和表型进行了表征。从地理上不同的HIV-1分离株中选择环境克隆，这些克隆对中和没有异常敏感或抗性，并且包含广泛的遗传、抗原和地理多样性。这些参考试剂将促进能力测试和其他旨在提高各实验室测定性能的验证工作，并可用于疫苗诱导的中和抗体的标准化评估。

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.