Studies of IGHV Germline Gene Polymorphism, Utilization & Shifting for Seasonal Influenza Vaccine Induced Broadly Neutralizing Antibody Responses

IGHV种系基因多态性研究及利用

• 批准号: 9009117
• 负责人: Wayne A. Marasco
• 金额: $ 75.64万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-11-09 至 2020-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9009117
• 关键词: Adult; Affect; Alleles; Antibodies; Antibody Formation; Antibody Repertoire; Antibody Response; Antigen Targeting; B-Lymphocytes; Binding; Biological; Blood specimen; Clinical Data; Cloning; Copy Number Polymorphism; Dana-Farber Cancer Institute; Data; Enrollment; Epitopes; Family; Frequencies; Genes; Genetic Polymorphism; Genetic Variation; Genotype; Goals; Health Personnel; Health Status; Heating; Hemagglutinin; Heterogeneity; Human; Immune; Immune Response Genes; Immune system; Immunity; Immunoglobulin M; Immunoglobulins; Individual; Influenza; Influenza A Virus, H5N1 Subtype; Influenza A virus; Influenza Hemagglutinin; Influenza vaccination; Maps; Measures; Membrane Glycoproteins; Memory; Memory B-Lymphocyte; Monitor; Outcome; Phenotype; Plasmablast; Play; Population; Recording of previous events; Recruitment Activity; Research Personnel; Role; ST14 gene; Sampling; Serological; Serum; Shapes; Structure; T-Lymphocyte; United States National Institutes of Health; Vaccination; Vaccine Design; Vaccines; Viral; anti-influenza; base; cohort; combinatorial; cytokine; design; genetic linkage; health difference; human monoclonal antibodies; influenza virus vaccine; inter-individual variation; interest; neutralizing antibody; neutralizing vaccine; pathogen; personalized medicine; public health relevance; response; seasonal influenza; stem; vaccine response; vaccine trial

 DESCRIPTION (provided by applicant): Neutralizing antibody responses to influenza infection and vaccination is highly variable in the population which can be explained by differences in health status, exposure history, and polymorphism of immune response genes. Since protection is also correlated with neutralizing antibody titers, any role that immunoglobulin germline gene polymorphism may play in this variability is important to establish, but has been difficult to investigate due to the use of numerous V, D and J genes in the genesis of immunoglobulins and the enormous combinatorial diversity that results from the pairing of rearranged VH and VL genes. However, the discovery of biased usage of the IGHV1-69 germline gene in antibodies targeting the stem domain of (HA) hemagglutinin (HV1-69-sBnAbs) has provided a unique opportunity to investigate any biological role that IGHV1-69 germline gene polymorphism might have in the anti-influenza antibody response. Importantly, HV1-69-sBnAbs mostly originate from the IGHV1-69 51p1 allele-like group that encodes a conserved CDR-H2 Phe54 that serves as major anchor residue with HA. Here we present an association analysis between IGHV1-69 polymorphism and the frequency of stem reactive Abs by using blood samples and clinical data obtained from an NIH H5VN04 vaccine trial. Our data shows a remarkable effect of IGHV1-69 locus polymorphism on the serum BnAb titers to the HA stem. Analyses of their expressed Ab repertoires have also provided evidence that copy number variation (CNV) also impacts IgM HV1-69-sBnAb responses. Results of heat map analysis show that there may be a shift in global IGHV germline gene usage in individuals based on the IGVH1-69 genotype. We propose that underlying genetic variation within the human IGHV1-69 locus might explain a significant fraction of inter- individual variation to influenza vaccination. Here we plan to enroll a cohort o 100 adult healthcare workers to participate in a study to examine the effects of IGHV1-69 polymorphism in shaping the BnAb response to seasonal influenza vaccination. Three blood specimens will be obtained from each subject, pre-vaccination, day 7 post vaccination (plasmablast peak) and at 32-56 days post-vaccination (memory B cell pool). In Aim 1 pre vaccination blood samples will be used to study the role IGHV1-69 locus polymorphism in shaping global IGHV germline gene utilization and in establishing the anti-influenza BnAb memory B cell pool. Genotyping will be by SNP and copy number variation (CNV) and phenotyping by serologic MN, HAI, HA binding studies, anti- HA single B cell cloning and antibodyome analysis. In Aim 2 heterogeneity at the IGHV1-69 locus will effect global germline gene utilization in the BnAb response to seasonal influenza vaccination. We propose that there is a direct genotypic-phenotypic relationship at the IGHV1-69 locus with regard to an individual's capacity to mount an anti-influenza BnAb response. This study is designed to provide the first evidence of this linkage. This new information on the role of IGHV polymorphism in influenza immunity can be used translationally to predict and monitor vaccine responses and to design vaccines to achieve universal responsiveness.

 描述（由申请人提供）：针对流感感染和疫苗接种的中和抗体反应在人群中差异很大，这可以通过健康状况、暴露史和免疫反应基因多态性的差异来解释。由于保护也与中和抗体滴度相关，因此确定免疫球蛋白种系基因多态性在这种变异中可能发挥的任何作用很重要，但由于在免疫球蛋白的发生中使用了大量的V、D和J基因以及重排的VH和VL基因配对产生的巨大组合多样性，因此很难研究。然而，发现 IGHV1-69 种系基因在针对 (HA) 血凝素干结构域 (HV1-69-sBnAbs) 的干结构域的抗体中存在偏差，为研究 IGHV1-69 种系基因多态性在抗流感抗体反应中可能发挥的生物学作用提供了独特的机会。重要的是，HV1-69-sBnAb 主要源自 IGHV1-69 51p1 等位基因样组，编码保守的 CDR-H2 Phe54，充当 HA 的主要锚定残基。在这里，我们利用 NIH H5VN04 疫苗试验获得的血液样本和临床数据，对 IGHV1-69 多态性与干反应性抗体频率之间的关联分析。我们的数据显示 IGHV1-69 位点多态性对 HA 干的血清 BnAb 滴度有显着影响。对它们表达的抗体库的分析也提供了证据，证明拷贝数变异 (CNV) 也会影响 IgM HV1-69-sBnAb 反应。热图分析结果表明，基于 IGVH1-69 基因型的个体中全球 IGHV 种系基因的使用可能发生变化。我们认为，人类 IGHV1-69 基因座内的潜在遗传变异可能可以解释流感疫苗接种的个体间变异的很大一部分。在这里，我们计划招募 100 名成年医护人员参加一项研究，以检查 IGHV1-69 多态性在影响 BnAb 对季节性流感疫苗接种反应中的影响。将从每个受试者采集三个血液样本，分别是疫苗接种前、疫苗接种后第 7 天（浆母细胞峰值）和疫苗接种后 32-56 天（记忆 B 细胞池）。在目标 1 中，将使用疫苗接种前的血液样本来研究 IGHV1-69 位点多态性在塑造全球 IGHV 种系基因利用和建立抗流感 BnAb 记忆 B 细胞库中的作用。基因分型将通过 SNP 和拷贝数变异 (CNV) 进行，表型分型则通过血清学 MN、HAI、HA 结合研究、抗 HA 单 B 细胞克隆和抗体组分析进行。在目标 2 中，IGHV1-69 位点的异质性将影响 BnAb 对季节性流感疫苗接种反应中的整体种系基因利用。我们认为 IGHV1-69 基因座与个体产生抗流感 BnAb 反应的能力存在直接的基因型-表型关系。本研究旨在提供这种联系的第一个证据。关于 IGHV 多态性在流感免疫中的作用的这一新信息可用于预测和监测疫苗反应，并设计疫苗以实现普遍反应。