ANTIBODY NEUTRALIZATION AND ESCAPE IN SUBTYPE C HIV-1

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 HIV-1感染中的中和抗体针对包膜糖蛋白gp120和gp41。然而，HIV-1Env利用高效但不明确的机制来逃避抗体介导的中和。我们缺乏关于B细胞反应、自体中和靶点以及C亚型感染中病毒逃逸机制的信息，这表明我们对世界上最主要的HIV-1变异株的理解存在巨大差距。这些研究的目标仍然是确定B细胞靶标，并使用一套独特的试剂，在新感染的C亚型患者中确定病毒中和和逃逸的机制，这些试剂来自赞比亚队列中新感染的C亚型HIV-1感染者。 在本报告所述期间，该项目又延长了五年的资金。在此期间，我们在两名C亚型HIV-1感染的赞比亚受试者中，以前所未有的详细描述了最初对感染病毒的自体NAB反应以及随后的病毒逃逸途径。中和抗体最初针对表面暴露的包膜高变域，病毒使用多个分子途径和补偿机制来逃避中和，即使在单个患者中也是如此。我们已经从其中一个赞比亚人身上产生了两个产生B细胞杂交瘤的单抗，并绘制了自体中和靶点和病毒逃逸突变的图谱。 我们已经从这个受试者身上收集了额外的可存活的PBMC，以分离具有不同特异性的单抗。我们收集了赞比亚另外10名受试者的纵向血浆和可见冷冻的PBMC样本，用于扩大中和和逃逸的研究，并产生更多产生自体中和单抗的B细胞杂交瘤。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Neutralizing antibodies in HIV-1 infection are directed against the envelope (Env) glycoproteins gp120 and gp41. However, HIV-1 Env utilizes highly effective, but poorly defined, mechanisms to evade antibody-mediated neutralization. Our lack of information about the B cell responses, targets of autologous neutralization, and mechanisms of viral escape in subtype C infection represents a significant gap in our understanding of the most predominant HIV-1 variants worldwide. The objective of these studies continues to be identifying B cell targets and defining mechanisms of virus neutralization and escape in newly infected subtype C patients using a unique set of reagents derived from newly subtype C HIV-1 infected subjects in a Zambian cohort. This project was renewed for an additional five years of funding during the reporting period. During this period, we have characterized the initial autologous Nab response against the infecting virus, and the viral escape pathways that ensue, in unprecedented detail in two subtype C HIV-1 infected Zambian subjects. Neutralizing antibodies initially target the surface exposed hyper-variable domains of the envelope, and the virus uses multiple molecular pathways and compensatory mechanisms to escape from neutralization, even within a single patient. We have generated two monoclonal antibody producing B cell hybridomas from one of these Zambian subjects, and mapped the autologous neutralization targets and viral escape mutations. We have collected additional viable PBMC from this subject to isolate monoclonal antibodies with different specificities. We have collected longitudinal plasma and viably frozen PBMC samples from 10 additional subjects in Zambia for expanded studies of neutralization and escape, and generation of more B cell hybridomas that produce autologous neutralizing monoclonal antibodies.