Characterization of Novel Polyreactive Anit-HIV Anitbodies Autoimmunity

描述(由申请人提供)： 一种有效的艾滋病毒疫苗应该在不同的人群中诱导针对大多数或所有艾滋病毒亚型的广泛中和抗艾滋病毒抗体[bnHIV-Ab]。尽管通过接种疫苗诱导针对艾滋病毒的中和抗体已被证明是一个挑战，但患有自身免疫性疾病的人经常产生多反应抗体，也能中和艾滋病毒感染。这项研究的长期目标是开发能够诱导广泛反应的中和抗体的策略，以防止被各种临床相关菌株感染。假设患有自身免疫的青少年，无论是否同时感染艾滋病毒，都可能拥有独特的外周B细胞，这些B细胞产生具有VH CDR3区的多反应抗体，这是已知的bnHIV-ab的特征。实现研究目标的策略使用探索性/发展性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细胞谱系的深度，通过创建新的噬菌体展示文库来增加可用的广泛中和艾滋病毒-1抗体的范围，并鉴定产生这些抗体的B细胞亚群[S]。研究人群包括一组独特的患有自身免疫的艾滋病毒感染青少年，以及患有自身免疫或艾滋病毒感染的个人。提出了两个相关的具体目标：1.寻找产生具有长VH CDR3的广谱反应性中和抗体的特定B细胞类型；2.开发和鉴定广谱中和HIV抗体。具体目标1将通过多色流式细胞术确定外周血B细胞群的特征，通过高通量454生命科学焦磷酸测序VH CDR3结构域来检测受试者IgG和IgM抗体库的深度，并确定长长的疏水性VH CDR3结构域在B细胞亚群中的定位。特异性目标2将抗体噬菌体展示的能力与自身免疫个体相结合，后者富含产生多反应自身抗体的B细胞，以开发一个定制的文库，该文库将针对复杂的病毒亚型混合物进行筛选，以分离出bnHIV-Ab。这项拟议的研究将提供有关人类免疫的细胞方面和抗体库的分子多样性的重要基础信息，并导致产生具有新的特异性和/或生物化学特征的广泛中和HIV抗体。艾滋病毒/艾滋病是一种全球流行病，目前没有疫苗，也没有治愈方法。尽管通过接种疫苗诱导针对艾滋病毒的中和抗体已被证明是一个巨大的挑战，但患有自身免疫性疾病的人经常制造也能中和艾滋病毒感染的抗体。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。这些结果将在理解对艾滋病毒的免疫反应方面取得重大进展，并将形成开发新的疫苗策略以诱导有效的抗艾滋病毒反应的基础。

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.