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

 描述(由申请人提供)：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的多态来解释。由于保护也与中和抗体效价相关，因此免疫球蛋白胚系基因多态性在这种变异性中可能扮演的任何角色都是重要的，但由于在免疫球蛋白的起源中使用了大量的V、D和J基因，以及由重排的VH和VL基因配对产生的巨大组合多样性，因此很难进行研究。然而，IGHV1-69胚系基因在针对血凝素(HA)干区(HV1-69-sBnAbs)的抗体中的偏向使用的发现为研究IGHV1-69胚系基因多态性在抗流感抗体应答中的生物学作用提供了一个独特的机会。重要的是，HV1-69-sBnAbs主要来源于IGHV1-69 51p1等位基因群，编码一个保守的CDR-H2 Phe54，作为HA的主要锚定残基。在这里，我们通过使用血液样本和来自NIH H5VN04疫苗试验的临床数据，对IGHV1-69多态与干细胞反应性抗体的频率进行了关联分析。我们的数据显示IGHV1-69基因座的多态对HA干的血清bNab滴度有显著的影响。对其表达的抗体谱系的分析也提供了拷贝数变异(CNV)也影响IgMHv1-69-sBnAb反应的证据。热图分析结果表明，基于IGVH1-69基因的个体的全球IGHV生殖系基因使用可能发生了变化。我们认为，人类IGHV1-69基因座的潜在遗传变异可能解释了流感疫苗接种的很大一部分个体间变异。在这里，我们计划招募100名成人医护人员参加一项研究，以检验IGHV1-69基因多态在影响bNab对季节性流感疫苗接种的反应中的作用。每个受试者将在接种前、接种后第7天(浆母细胞峰值)和接种后32-56天(记忆B细胞池)采集三份血液样本。目的1采用免疫前血样研究IGHV1-69基因座多态性在全球IGHV种系基因利用和建立抗流感病毒bNab记忆B细胞库中的作用。将通过SNP和拷贝数变异(CNV)进行基因分型，并通过血清学MN、HAI、HA结合研究、抗HA单个B细胞克隆和抗体分析进行表型分析。在AIM 2中，IGHV1-69基因座的异质性将影响全球生殖系基因在bNab对季节性流感疫苗接种的反应中的利用。我们认为IGHV1-69基因座与个体启动抗流感bNab反应的能力有直接的基因-表型关系。这项研究旨在提供这种联系的第一个证据。这一关于IGHV多态在流感免疫中作用的新信息可用于预测和监测疫苗应答，并设计疫苗以实现普遍应答。

 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.