Characterization of Novel Polyreactive Anit-HIV Anitbodies Autoimmunity

描述（由申请人提供）： 有效的HIV疫苗应在不同人群中诱导针对大多数或所有HIV亚型的广泛中和抗HIV抗体[bnHIV-Ab]。尽管通过疫苗接种诱导针对艾滋病毒的中和抗体已被证明是一项挑战，但患有自身免疫性疾病的个体经常产生多反应性抗体，这些抗体也能中和艾滋病毒感染。这项研究的长期目标是开发诱导广泛反应性中和抗体的策略，这些抗体可以预防各种临床相关菌株的感染。该假说认为，伴有或不伴有HIV感染的自身免疫性青少年可能具有独特的外周B细胞，该细胞产生具有长且疏水的VH CDR 3区的多反应性抗体，这是已知bnHIV-ab的特征。实现研究目标的策略使用探索性/开发性R21机制和跨学科方法，应用高影响力的基础免疫学研究和创新方法来探索B细胞库的深度，通过创建新型噬菌体展示文库来增加广泛中和HIV-1抗体的面板，并鉴定产生这些抗体的B细胞亚群。研究人群包括一组独特的自身免疫性HIV感染青少年，以及自身免疫性或HIV感染的个体。提出了两个相关的具体目标：1。寻找产生具有长VH CDR 3的宽反应性中和抗体的特异性B细胞类型;和2.开发和鉴定广泛中和的HIV抗体。特异性目标1将通过多色流式细胞术表征外周血B细胞群，通过VH CDR 3结构域的高通量454 Life Science焦磷酸测序检查受试者中IgG和IgM抗体库的深度，并确定长疏水性VH CDR 3结构域在B细胞亚群中的定位。Specific Aim 2将抗体噬菌体展示的能力与自身免疫个体相结合，这些个体富含产生多反应性自身抗体的B细胞，以开发定制文库，该文库将针对病毒体亚型的复杂混合物进行筛选以分离bnHIV-Ab。拟议的研究将提供有关人类免疫细胞方面和抗体库分子多样性的重要基础信息，并导致产生具有新特异性和/或生化特征的广泛中和HIV抗体。艾滋病毒/艾滋病是一种全球性流行病，目前没有疫苗，也没有治愈方法。虽然通过接种疫苗诱导抗HIV的中和抗体已被证明是一个重大挑战，但患有自身免疫性疾病的个体经常产生也中和HIV感染的抗体。拟议的探索性/发展性研究将应用最先进的技术，对自身免疫个体的艾滋病毒特异性抗体反应进行全面的细胞和分子分析。这些结果将为理解对艾滋病毒的免疫反应提供重大进展，并将为开发新的疫苗策略以诱导有效的抗艾滋病毒反应奠定基础。

DESCRIPTION (provided by applicant): An effective HIV vaccine should induce in a diverse human population broadly-neutralizing anti-HIV antibodies [bnHIV-Ab] that target most or all HIV subtypes. Although inducing neutralizing antibodies against HIV by vaccination has proven to be a challenge, individuals with autoimmune disease frequently make polyreactive antibodies that also neutralize HIV infection. The long-term goal for the research proposed is to develop strategies that lead to induction of broadly reactive neutralizing antibodies that can prevent infection by a wide variety of clinically relevant strains. The hypothesis is that adolescents with autoimmunity with or without concurrent HIV infection are likely to harbor unique peripheral B cells that produce polyreactive antibodies with long and hydrophobic VH CDR3 regions, which are characteristics of known bnHIV-ab. The strategy to accomplish the research goals uses the exploratory/developmental R21 mechanism and an interdisciplinary approach to apply high impact basic immunology studies and innovative methods to explore the depth of the B-cell repertoire, to increase the panel of broadly neutralizing HIV-1 antibodies available by creating a novel phage display library, and to identify the B cell subset[s] that produce these antibodies. The study population includes a unique group of HIV infected adolescents with autoimmunity, as well as individuals with autoimmunity or HIV infection. Two related Specific Aims are proposed: 1. to search for specific B cell types producing broad reactive neutralizing antibodies with long VH CDR3; and 2. to develop and characterize broadly-neutralizing HIV antibodies. Specific Aim 1 will characterize peripheral blood B cell populations by multi-color flow cytometry, examine the depth of IgG and IgM antibody repertoires in subjects by high-throughput 454 Life Science pyrosequencing of VH CDR3 domains, and determine the localization of long, hydrophobic VH CDR3 domains within subsets of B cells. Specific Aim 2 combines the power of antibody phage display with autoimmune individuals, who are enriched for B cells that produced polyreactive autoantibodies, to develop a custom library that will be screened against a complex mixture of virion subtypes to isolate bnHIV-Ab. The proposed research will provide significant fundamental information about cellular aspects of human immunity and the molecular diversity of the antibody repertoire, and result in generation of broadly neutralizing HIV antibodies with novel specificities and/or biochemical characteristics. HIV/AIDS is a global pandemic for which there is currently no vaccine and no cure. Although inducing neutralizing antibodies against HIV by vaccination has proven to be a significant challenge, individuals with autoimmune disease frequently make antibodies that also neutralize HIV infection. The exploratory/developmental research proposed will apply state-of-the-art technology for a comprehensive cellular and molecular analysis of HIV-specific antibody responses in autoimmune individuals. The results will provide major advancements in understanding the immune response to HIV and will form a basis for developing novel vaccine strategies to induce an effective anti-HIV response.