Characterizing HIV-1 Envs associated with potent bnAbs agains QNEs

表征与针对 QNE 的有效 bnAb 相关的 HIV-1 Envs

• 批准号: 7904631
• 负责人: Lynn Morris
• 金额: $ 37.21万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7904631
• 关键词: Amino Acids; Animal Model; Antibodies; Antibody Specificity; Antigens; Autologous; Biological Assay; Cercopithecine Herpesvirus 1; Collaborations; Data; Development; Epitopes; Evolution; Genes; Genetic Variation; HIV; HIV Envelope Protein gp120; HIV vaccine; HIV-1; Individual; Infection; Maps; Methods; Modeling; Monoclonal Antibodies; Mutation; Neutralization Tests; Participant; Peptides; Population; Positioning Attribute; Recombinants; Resistance; Role; Sampling; Serum; Site; Site-Directed Mutagenesis; Specificity; Structure; Testing; Time; Vaccine Research; Variant; Viral; Virus; cohort; design; interest; mutant; neutralizing antibody; novel; research study

The aim of this project is to identify and characterize HIV-infected individuals from the CAPRISA 002 cohort with broadly neutralizing antibodies that target quaternary neutralization epitopes (QNE) on the HIV envelope. To date we have identified 2 such individuals from among the 28 so far studied (Dr Morris). One of these, CAP256 developed potent antibodies against QNE that included the V1V2 region (Dr Moore). Interestingly this individual was super-infected (Dr Williamson) and viral evolution studies will be performed to indentify the second infecting virus as well as the recombinant strains to ascertain their role in the development of these antibodies. As part of this project we anticipate identifying another 9 subjects with neutralizing antibodies that target QNE from among the additional 90 that will be screened. This will be done by performing neutralization assays on a large panel of multi-subtype pseudoviruses; those with >60% neutralization breadth and who do not have anti-gp120 or anti-MPER neutralizing antibodies will be considered as having antibodies against QNE. Envelope genes from selected time-points during the development of neutralization breadth will be sequenced and functional pseudotypes tested for neutralization sensitivity to identify putative sites. We will furthermore make use of chimeric and mutant viruses to identify neutralization escape mutations as a way of identifying the antibody targets. The fine mapping and structure of these QNE epitopes will be done in collaboration with Dr Pinter (Project 1). Monoclonal antibodies will be made from those individuals where the neutralizing activity is attributable to a single specificity by Dr James Robinson (Project 4). This information will be used to design immunogens that will be tested in animal models by Dr Shiu-lok (Project 3).

该项目的目的是从 CAPRISA 002 队列中识别和表征 HIV 感染者，其具有针对 HIV 包膜上四元中和表位 (QNE) 的广泛中和抗体。迄今为止，我们已从迄今为止研究的 28 人中识别出 2 人（莫里斯博士）。其中之一，CAP256 开发了针对 QNE 的有效抗体，其中包括 V1V2 区域（Moore 博士）。 有趣的是，这个人被超级感染（威廉姆森博士），将进行病毒进化研究，以鉴定第二种感染病毒以及重组毒株，以确定它们在这些抗体发展中的作用。作为该项目的一部分，我们预计从将要筛选的另外 90 名受试者中鉴定出另外 9 名具有针对 QNE 的中和抗体的受试者。这将通过对大量多亚型假病毒进行中和测定来完成；中和广度 >60% 且不具有抗 gp120 或抗 MPER 中和抗体的患者将被视为具有抗 QNE 抗体。对中和宽度发展过程中选定时间点的包膜基因进行测序，并测试功能假型的中和敏感性，以识别推定位点。我们还将进一步利用嵌合病毒和突变病毒来识别中和逃逸突变，作为识别抗体靶点的一种方式。这些 QNE 表位的精细绘图和结构将与 Pinter 博士合作完成（项目 1）。 James Robinson 博士（项目 4）将从中和活性归因于单一特异性的个体中制备单克隆抗体。这些信息将用于设计免疫原，并由 Shiu-lok 博士（项目 3）在动物模型中进行测试。