Influenza Immunity: Protective Mechanisms Against Pandemic Respiratory Virus

描述（由申请人提供）：在这项名为“流感免疫：对抗大流行性呼吸道病毒的保护机制”的续期申请中，我们打算在上一个资助期取得进展的基础上再接再厉，并牢记两个总体目标：第一个目标是在不同年龄组和不同的疫苗模式中，更广泛和深入地扩展我们对流感疫苗的人体免疫应答的知识并利用同卵双胞胎和异卵双胞胎来探讨遗传学对这些反应的影响。我们正在追求的第二个新主题是利用我们对两种许可的流感疫苗的反应的分析来建立一个独特的人类免疫学数据集和“指标”，我们可以开始识别生物标志物，以确定什么是健康和有效的反应，什么不是。在我们重新组建的团队中，我们在免疫学、传染病、疫苗学、分子生物学、免疫监测、生物工程、遗传学和生物信息学方面具有显著优势。我们也有一个杰出的临床核心，在招募和处理所有年龄段的受试者进入我们的疫苗研究方面非常出色。加强我们的临床队列以及与SRI国际的新关系，SRI国际在先天性和适应性免疫方面有一个庞大而不断扩大的双胞胎登记处，其成员将构成我们学科基础的核心。为了从每个临床标本中提取最大量的信息，我们在基础设施中增加了两个新的核心--斯坦福大学人类免疫监测核心（Human Immune Monitoring Core）和生物信息学核心（Bioinformatics Core），前者提供对每个受试者的深入和统一的免疫学分析，后者组织大量数据并帮助研究人员进行解释。我们专注于B和T细胞反应和库以及生物标志物和信号通路，在我们的技术开发项目中，我们寻求突破剧目和单细胞分析的界限，并开发创新的新生物信息学工具，这对理解免疫系统的复杂性和对流感病毒的反应。 相关性（参见说明）：流感病毒是一个严重的公共卫生问题，不仅因为它引起的疾病和死亡，而且因为它可能以大流行病或生物恐怖主义武器的形式造成更大的伤害。在本更新申请中，我们寻求更好地了解我们目前的流感疫苗如何在与人类免疫系统的相互作用方面发挥作用，以及如何利用这种相互作用来定义免疫健康的“指标”。 项目1：B细胞对流感的免疫力（格林伯格，H） 项目1描述（由申请人提供）：流感疫苗开发中一个艰巨且几乎独特的挑战是，人群反复暴露于不断变化的流感病毒，这使得先前具有免疫力的个体易受新出现的毒株的攻击，从而导致流感的年度流行甚至大流行。虽然人类免疫系统具有以其庞大的抗体库应对高度多样化的病毒抗原的内在能力，（估计为1011的量级），我们假设这种潜在的多样性在某些年龄组中受到显著限制，例如老年人和非常年轻的人，并且在某些年龄组中接种某些类型的疫苗后没有完全实现，例如，与幼儿中的减毒活流感疫苗（LAIV）相比，三价灭活流感疫苗（TIV）^在本申请中，我们建议。使用血清学和分子学方法解决这些问题，重点是最易受流感发病率和死亡率影响的年龄组-幼儿和老年人。该提案的具体目标是：A1。比较LAIV或TIV免疫后针对疫苗株（同型反应性）和错配病毒株（异源变异反应性）的抗体应答。我们将描述和比较接种这两种流感疫苗的幼儿、健康成人和老年人的配对血清样本中的异源变异抗体应答。我们还将克隆和表达免疫球蛋白（IG）基因，从个别抗体分泌细胞（ASC）后不久，在不同年龄组的TIV或LAIV免疫接种，并比较这些单克隆抗体的特异性和亲和力对同型和异型流感病毒。A2.为了确定影响编码流感特异性抗体的IG基因序列的因素，并确定特异性IG基因序列使用与功能之间的关系，我们将对用LAIV或TIV免疫后从ASC中分离的IG基因序列进行系统分析，并将IG基因的序列特征与针对不同流感毒株的抗体反应性相关联。特别是，我们将比较老年人，年轻人和儿童之间的IG基因序列，LAIV和TIV的收件人之间和IgG和伊加同种型之间。 相关性（参见说明）：本研究与流感疫苗的设计和给药直接相关，用于预防大流行性流感和流行性流感，特别是在非常年幼的儿童和老年人中。它还将提供重要的信息，有关机制负责产生保护性B细胞免疫力，对一系列其他重要的人类病原体和生物恐怖主义制剂。

DESCRIPTION (provided by applicant): In this renewal application entitled "influenza Immunity: Protective Mechanisms Against Pandemic Respiratory Virus," we intend to build on the progress we have made in the previous granting period with two overall objectives in mind: the first is to extend our knowledge of the human immune response to influenza vaccine much more broadly and deeply across different age groups and with different vaccine modalities and to probe the influence of genetics on these responses using monozygotic and dizygotic twin pairs. A second and new theme that we are pursuing is to use our analyses of responses to the two licensed influenza vaccines to establish a unique dataset and "metrics" of human immunology in which we can begin to identify biomarkers for what is a healthy and effective response versus what is not. In our reformulated team, we have significant strengths in immunology, infectious diseases, vaccinology, molecular biology, immune monitoring, bioengineering, genetics and bioinformatics. We also have an outstanding clinical core that has been extremely good at recruiting and processing subjects of all ages into our vaccine studies. Enhancing our clinical cohort as well as a new relationship with SRI International, which has a large and expanding twin registry in innate and adaptive immunity, the members of which will form the core of our subject base. In order to extract the maximum amount of information from each clinical specimen, we have added two new cores to our infrastructure-the Stanford Human Immune Monitoring Core to provide an in depth and uniform immunological analysis of each subject, and a Bioinformatics Core to organize the large quantities of data and to help investigators interpret it. In our projects, we focus on B and T cell responses and repertoire as well as biomarkers and signaling pathways, and in our Technology Development projects, we seek to push the boundaries of repertoire and single cell analysis and also to develop innovative new bioinformatics tools that will be vital to understanding the complexities of the immune system and the response to the influenza virus. RELEVANCE (See instructions): The Influenza virus represents a serious public health concern not just because of the illness and death that it causes but also in its potential for much greater harm in the form of a pandemic or a weapon of bioterrorism. In this renewal application, we seek to better understand how our current influenza vaccines work in terms of their interactions with the human immune system and how that interaction can be utilized to define "metrics" of immunological health. PROJECT 1: B Cell Immunity to Influenza (GREENBERG, H) PROJECT 1 DESCRIPTION (provided by applicant): A formidable and virtually unique challenge in influenza vaccine development is that the human population is repeatedly exposed to changing influenza viruses, which render previously immune individuals vulnerable to newly emerged strains, thus causing annual epidemics or even pandemics of influenza. While the human immune system has the intrinsic capability of coping with highly diversified viral antigens with its enormous antibody repertoire (estimated at a magnitude of 1011), it is our hypothesis that this potential diversity is significantly restricted in certain age groups, such as the elderly and the very young, and is not fully realized following immunization with certain types of vaccine in some age groups, such as the trivalent inactivated influenza vaccine (TIV) compared to the live attenuated influenza vaccine (LAIV) in young children^ In this application we propose to. address these, issues using, both serologic and molecular approaches, with an emphasis on the age groups most vulnerable to influenza morbidity and mortality - very young children and the elderly. The specific aims of the proposal are: A1. To compare the antibody responses against the vaccine strains (homotypic reactivity) and mismatched virus strains (heterovariant reactivity) after immunization with LAIV or TIV. We will characterize and compare the heterovariant antibody responses in a collection of paired serum samples from young children, healthy adults and elderly who are vaccinated with these two types of influenza vaccines. We will also clone and express immunoglobulin (Ig) genes from individual antibody-secreting cells (ASCs) elicited shortly after immunization with TIV or LAIV in the different age groups and compare the specificity and affinity of these monoclonal antibodies against both homotypic and heterovariant influenza viruses. A2. To identify factors affecting the sequences of Ig genes encoding influenza-specific antibodies and determine the relationship between specific Ig gene sequence usage and function, we will carry out a systematic analysis of the sequences of Ig genes isolated from ASCs after immunization with LAIV or TIV, and relate the sequence characteristics of Ig genes to antibody reactivity against different influenza strains. In particular, we will compare the Ig gene sequences between the elderly, younger adults and children, between the recipients of LAIV and TIV and between IgG and IgA isotypes. RELEVANCE (See instructions): This study is directly relevant to the design and administration of influenza vaccines for the prevention of pandemic and epidemic influenza, especially in the very young children and the elderly. It will also provide important information regarding the mechanisms responsible for the generation of protective B cell immunity against a wide array of other important human pathogens and bioterrorism agents.