Measurement of HIV Neutralization Using Quantitative Assays

使用定量测定测量 HIV 中和作用

• 批准号: 8556086
• 负责人: John Mascola
• 金额: $ 22.05万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8556086
• 关键词: Animals; Antigens; Biological Assay; Cells; Clinical; Genetic; Goals; Guidelines; HIV; HIV Envelope Protein gp120; HIV-1; Human; Immune; Immune response; Immunity; Individual; Laboratories; Length; Luciferases; Measurement; Measures; Monoclonal Antibodies; Performance; Phenotype; Plasmids; Process; Reagent; Recombinants; Reporter Genes; Research Personnel; Resistance; Sampling; Serum; Standardization; Testing; Vaccines; Validation; Viral; Virus; base; env Glycoproteins; good laboratory practice; gp160; high throughput screening; improved; interest; neutralizing antibody; plasmid DNA; vaccine candidate; vaccine evaluation; virology

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.

在JC53-BL（TZM-BL）细胞中的荧光素酶报告基因测定中，将单个GP160克隆作为Env-Pseudotyped病毒进行筛查。 对功能性ENV克隆进行了测序，并通过使用可溶性CD4，来自感染个体的单克隆抗体和血清样品的中和表型进行了特征，而重组GP120疫苗的非感染者和未感染的受体。 从地理上不同的HIV-1分离株中选择了不异常敏感或对中和的抗性，并包括广泛的遗传，抗原和地理多样性。 这些参考试剂将有助于熟练度测试和其他旨在改善实验室测定性能的验证工作，并可用于标准化疫苗引起的中和抗体的评估。 使用上面描述的参考病毒摄氏，BSL3核心病毒学实验室已经筛选了VRC研究人员产生的数百种动物血清。