Deep Sequencing to Identify B-Cell Precursors of HIV-1 Neutralizing Antibodies

深度测序鉴定 HIV-1 中和抗体的 B 细胞前体

• 批准号: 9086242
• 负责人: Xueling Wu
• 金额: $ 65.91万
• 依托单位: AARON DIAMOND AIDS RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9086242
• 关键词: Acute; Address; Affinity; Antibodies; Antibody Repertoire; Antibody Response; Antigens; Autologous; B-Lymphocytes; Binding; Blood; Blood Cells; Blood specimen; Cells; Cloning; Complex; Critical Pathways; Development; Epitopes; Genome; Glycoproteins; Goals; HIV-1; Health; Human; Immunity; Immunization; Individual; Infection; Macaca mulatta; Mining; Monoclonal Antibodies; Plasma; Process; Production; Receptors, Antigen, B-Cell; Resolution; Sampling; System; Technology; Testing; Time; Vaccination; Vaccine Design; Virus; antigen binding; deep sequencing; design; immunogenicity; insight; interest; lymph nodes; neutralizing antibody; neutralizing monoclonal antibodies; response; screening; simian human immunodeficiency virus; vaccine development

DESCRIPTION: Our recent breakthroughs in applying single B-cell probing and cloning technologies to isolate human antibodies capable of potently and broadly neutralizing HIV-1 primary isolates, along with others, have demonstrated the ability of human B-cell system to generate effective immunity against the virus. To progress on this encouraging discovery and unearth mechanisms by which broadly neutralizing antibodies (bnAbs) could be elicited by immunization, we are challenged by two unsolved and fundamental immunological questions: what are the naive and/or founder (n/f) B-cells and their B-cell receptor (BCR) sequences that must be immunologically selected to generate anti-HIV-1 bnAbs, and how do these selected n/f BCRs react with the HIV-1 envelope (Env) antigen? Having access to precious longitudinal samples from both SHIV-infected rhesus macaques (including lymph nodes) and HIV-1- infected seroconverters, we have a unique opportunity to address these questions by identifying the n/f BCRs responsible for bnAbs and determining their reactivity to the Env antigen. In addition to single B-cell analysis, we have also pioneered the application of deep sequencing technology to mine anti-HIV-1 bnAbs of the whole antibody repertoire. With the aid of these advanced technologies that offer high- resolution analysis of both single and high-number (millions) B-cells, it is feasible to track bnAb responses, thereby catapulting our quest to identiy bnAb-corresponding n/f BCRs. Furthermore, we will isolate the autologous infecting HIV-1 Envs and investigate their reactivity with the n/f BCRs of interest. We will test the hypotheses that n/ BCRs that give rise to bnAbs in SHIV-infected rhesus macaques and HIV-1-infected humans can be identified by deep sequencing of the subject antibodyome from longitudinally collected blood and lymph node cells, and that the n/f BCRs recognize the autologous infecting HIV-1 Envs. If successful, this project will delineate a complex and lengthy B-cell immunological process, revealing the much anticipated initial steps that the HIV-1 Env antigen must take to initiate an effective antibody response. This project will also generate a panel of bnAb-corresponding n/f BCRs that would be invaluable for immunogen screening and may also identify HIV-1 Envs that have the potential to prime these n/f BCRs.

描述：我们最近在应用单个B细胞探测和克隆技术方面的突破来分离能够有效，广泛中和的HIV-1主要分离株的人类抗体以及其他抗体，也证明了人类B细胞系统对病毒产生有效免疫的能力。 To progress on this encouraging discovery and unearth mechanisms by which broadly neutralizing antibodies (bnAbs) could be elicited by immunization, we are challenged by two unsolved and fundamental immunological questions: what are the naive and/or founder (n/f) B-cells and their B-cell receptor (BCR) sequences that must be immunologically selected to generate anti-HIV-1 bnAbs, and how do these selected N/F BCR与HIV-1信封（ENV）抗原反应？我们可以从SHIV感染的恒河猕猴（包括淋巴结）和HIV-1感染的血清抗体中获取宝贵的纵向样品，我们有一个独特的机会来通过识别负责BNABS的N/F BCR来解决这些问题，并确定其对Env antigen的反应性。除了单个B细胞分析外，我们还开创了深层测序技术在整个抗体库中的抗HIV-1 BNABs的应用。在这些高级技术的帮助下，它们对单个和高数字（数百万）b细胞进行高分辨率分析，跟踪BNAB响应是可行的，从而迅速降低了我们寻求识别bnab对应的n/f bcrs的追求。此外，我们将隔离自体感染HIV-1 ENV，并研究其对N/F BCR的反应性。我们将测试以下假设，即可以通过对纵向收集的血液和淋巴结细胞的受试者抗体细胞的深度测序来鉴定出SHIV感染的恒河猕猴和HIV-1感染的人的N/ BCR，并可以鉴定出N/ F BCR的N/ F BCR识别N/ F BCR识别自动志出的HIV-HIV-1 Envs。如果成功的话，该项目将描绘出复杂而冗长的B细胞免疫学过程，揭示HIV-1 Env抗原启动有效抗体反应所需的备受期待的初始步骤。该项目还将生成一组BNAB对应的N/F BCR，这对于免疫原筛查是无价的，并且还可以鉴定有可能启用这些N/F BCR的HIV-1 ENV。