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

 描述（由申请方提供）：人群中对流感感染和疫苗接种的中和抗体应答具有高度可变性，这可以通过健康状况、暴露史和免疫应答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，因此免疫球蛋白种系基因多态性在这种变异性中可能发挥的任何作用都很重要，但由于在免疫球蛋白的发生中使用了许多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，因此难以研究。然而，在靶向（HA）血凝素干域（HV 1 -69-sBnAb）的抗体中发现IGHV 1 -69种系基因的偏倚使用，为研究IGHV 1 -69种系基因多态性在抗流感抗体应答中可能具有的任何生物学作用提供了独特的机会。重要的是，HV 1 -69-sBnAb主要来源于IGHV 1 -69 51 p1等位基因样组，其编码作为HA的主要锚残基的保守CDR-H2 Phe 54。在这里，我们提出了IGHV 1 -69多态性和干细胞反应性抗体的频率之间的关联分析，使用血液样本和临床数据从NIH H5 VN 04疫苗试验。我们的数据显示IGHV 1 -69位点多态性对HA干细胞的血清BnAb滴度有显著影响。对它们表达的Ab库的分析也提供了拷贝数变异（CNV）也影响IgM HV 1 -69-sBnAb应答的证据。热图分析的结果显示，基于IGVH 1 -69基因型，个体中的全球IGHV生殖系基因使用可能发生变化。我们提出，人类IGHV 1 -69基因座内潜在的遗传变异可能解释流感疫苗接种的个体间变异的显著部分。在此，我们计划招募100名成年医护人员参与一项研究，以检查IGHV 1 -69多态性在形成季节性流感疫苗接种的BnAb应答中的作用。在接种前、接种后第7天（浆母细胞峰）和接种后第32-56天（记忆B细胞池），从每例受试者中采集3份血液标本。在目标1中，将使用接种前血样研究IGHV 1 -69基因座多态性在形成全球IGHV生殖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，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表型分型。在目的2中，IGHV 1 -69基因座的异质性将影响BnAb对季节性流感疫苗接种应答中的总体生殖系基因利用。我们认为IGHV 1 -69位点与个体产生抗流感病毒BnAb应答的能力存在直接的基因型-表型关系。这项研究旨在提供这种联系的第一个证据。IGHV多态性在流感免疫中的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.