Innate-like BCR activity as a template for universal vaccination against influenza virus

项目总结/摘要 该提案描述了R 01资助的框架，该资助由哈佛大学助理教授丹尼尔·林伍德博士提供。 哈佛医学院和麻省理工学院和哈佛大学拉根研究所的教员，以及布莱斯博士 Chackerian是该大学分子遗传学和微生物学系的副教授 来自新墨西哥州医学院。他们的研究中心是B细胞受体（BCR）抗原识别 生物学（Lingwood）和病毒样颗粒（VLP）疫苗平台的定向进化（Chackerian）。一起 他们建议开发一种通用流感疫苗设计的新策略。大多数疫苗引发的抗体 对该病毒的应答主要是免疫显性脱靶、非中和活性。然而，在这方面， Lingwood博士最近的工作表明，人类BCR是使用抗体基因IGHV 1 -69组装的 具有V区编码（先天性）特异性的功能保守的网站的脆弱性，茎- 流感刺突蛋白血凝素（HA）表位和广谱中和抗体（bnAb）靶点 应答为了实验性地评估这种基因编码模板，以构建通用疫苗，Dr。 Lingwood已经培育出了转基因小鼠，其中抗体通过正常的人类VDJ发育 重组，但其中V区的使用限于IGHV 1 -69。初步数据显示IGHV 1 -69 使用本身重新集中抗体对茎表位的反应，Lingwood博士发现的一个特征是， 依赖于IGHV 1 -69的茎接触CDRH 2中的单个基因编码的氨基酸。至今 Lingwood博士将此应用于bnAb诱导，建议用合理的- 设计的流感HA茎的三聚体和纳米颗粒展示，其触发先天样茎-表位 通过重建的种系IGHV 1 -69 BCR进行信号传导。通过这些实验，林伍德博士将提供 合理疫苗设计的一个重大范式转变，即通过 基因编码的抗体应答的“激活”和“扩增”。为临床确定一条路径 Lingwood博士和Chackerian博士已经将RNA噬菌体肽展示和亲和力 获得对IGHV 1 -69种系的V区具有多价亲和力的VLP疫苗的选择技术 BCR本质上是一种V区特异性引物，用于选择性扩大先天性干细胞靶向活性，即 否则通常被人类BCR多样性稀释。选择性IGHV 1 -69预充将在 Trianni小鼠，最新的行业标准人源化小鼠疫苗模型。然后，V区域预充将是 用IGHV 1 -69接合HA干免疫原加强，以刺激现在免疫显性的先天性样HA 干感应抗体反应。利用bnAb诱导的遗传基础旨在克服 流感疫苗接种的传统方法，并与此资助机会的目的一致 和更广泛的NIH使命。

Project Summary / Abstract This proposal describes the framework of an R01 grant by Dr. Daniel Lingwood, an Assistant Professor at Harvard Medical School and faculty member of the Ragon Institute of MGH, MIT and Harvard, and Dr. Bryce Chackerian, an Associate Professor in the Department of Molecular Genetics & Microbiology at the University of New Mexico School of Medicine. Their research centers on B cell receptor (BCR) antigen recognition biology (Lingwood) and directed evolution of virus-like particle (VLP) vaccine platforms (Chackerian). Together they propose to develop a new strategy for universal influenza vaccine design. Most vaccine-elicited antibody responses to this virus are dominated by immunodominant off-target, non-neutralizing activities. However, recent work from Dr. Lingwood indicates that human BCRs assembled using the antibody gene IGHV1-69 possess V region-encoded (innate-like) specificity for a functionally conserved site of vulnerability, the stem- epitope of the influenza spike protein hemagglutinin (HA) and target of broadly neutralizing antibody (bnAb) responses. To experimentally evaluate this as a gene-encoded template for building a universal vaccine, Dr. Lingwood has generated transgenic mice in which antibody development proceeds via normal human VDJ recombination, but where V region use is constrained to IGHV1-69. Preliminary data indicate that IGHV1-69 usage itself refocuses the antibody response to the stem epitope, a feature that Dr. Lingwood finds is dependent on a single gene-encoded amino acid in the stem-contacting CDRH2 of IGHV1-69. To now transduce this into bnAb elicitation, Dr. Lingwood proposes to immunize his humanized mice with rationally- designed trimeric and nanoparticle displays of influenza HA stem which trigger innate-like stem-epitope signaling by the reconstituted germline IGHV1-69 BCR. Through these experiments Dr. Lingwood will provide a major paradigm shift in rational vaccine design, namely that broad protection may be generated through `activation' and `amplification' of gene-encoded antibody responses. To define a pathway for clinical development, Drs. Lingwood and Chackerian have applied RNA bacteriophage peptide display and affinity selection technology to derive a VLP vaccine with multivalent affinity to the V region of IGHV1-69 germline BCR, in essence a V region-specific primer to selectively expand innate-like stem targeting activity that is otherwise normally diluted by human BCR diversity. Selective IGHV1-69 priming will be evaluated in the Trianni mouse, the latest industry-standard humanized mouse vaccine model. V region priming will then be boosted with IGHV1-69-engaging HA stem immunogens, to stimulate a now immunodominant innate-like HA stem sensing antibody response. Exploiting a genetic basis for bnAb elicitation aims to overcome the failure of traditional approaches to influenza vaccination and is consistent with the purpose of this funding opportunity and broader mission of the NIH.