Influenza Immunity: Protective Mechanisms Against Pandemic Respiratory Virus

描述(申请人提供)：在题为“流感免疫：预防大流行性呼吸道病毒的保护机制”的续期申请中，我们打算在上一次授权期取得的进展的基础上，考虑到两个总体目标：第一是更广泛和更深入地扩展我们对不同年龄段和不同疫苗类型的人类对流感疫苗的免疫反应的知识，并使用同卵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，利用我们对两种获得许可的流感疫苗的反应的分析，建立一个独特的人类免疫学数据集和“指标”，在其中我们可以开始识别哪些是健康和有效的反应，哪些不是。在我们重新制定的团队中，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拥有显著优势。我们还拥有出色的临床核心，在招募和处理各个年龄段的受试者进入我们的疫苗研究方面一直非常出色。加强我们的临床队列以及与SRI国际的新关系，SRI国际在先天免疫和适应性免疫方面拥有庞大且不断扩大的孪生注册，其成员将构成我们的学科基础的核心。为了最大限度地从每个临床标本中提取信息，我们在我们的基础设施中增加了两个新的核心-斯坦福人类免疫监测核心，以提供对每个对象的深入和统一的免疫分析，以及一个生物信息学核心，用于组织大量数据并帮助研究人员解释这些数据。在我们的项目中，我们专注于B和T细胞反应和谱系以及生物标记物和信号通路，在我们的技术开发项目中，我们寻求突破谱系和单细胞分析的界限，并开发创新的新生物信息学工具，这些工具对于了解免疫系统的复杂性和对流感病毒的反应至关重要。 相关性(见说明)：流感病毒是一个严重的公共卫生问题，不仅是因为它造成的疾病和死亡，而且还因为它有可能以大流行或生物恐怖主义武器的形式造成更大的危害。在这一续期申请中，我们试图更好地了解我们目前的流感疫苗在与人类免疫系统的相互作用方面是如何发挥作用的，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(格林伯格，H) 项目1说明(申请人提供)：流感疫苗开发中一个艰巨且几乎独一无二的挑战是，人类反复接触不断变化的流感病毒，使以前免疫的个人容易受到新出现的毒株的影响，从而导致每年一次的流感流行甚至大流行。虽然人类免疫系统具有通过其庞大的抗体库(估计为1011个数量级)来应对高度多样化的病毒抗原的内在能力，但我们的假设是，这种潜在的多样性在某些年龄组中受到显著限制，并且在某些年龄组中接种某些类型的疫苗后不能完全实现，例如在幼儿中使用三价灭活流感疫苗(TIV)与减毒活流感疫苗(LAIV)进行比较。使用血清学和分子方法解决这些问题，重点放在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免疫后对疫苗株(同型反应性)和错配病毒株(异型反应性)的抗体反应。我们将对接种了这两种流感疫苗的幼儿、健康成年人和老年人的配对血清样本中的异型抗体反应进行表征和比较。我们还将从不同年龄组的TIV或LAIV免疫后不久诱导的单个抗体分泌细胞(ASCs)中克隆和表达免疫球蛋白(Ig)基因，并比较这些单抗对同种和异种流感病毒的特异性和亲和力。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，并确定特定的Ig基因序列的使用与功能的关系，我们将对LAIV或TIV免疫后的ASCs分离的Ig基因序列进行系统分析，并将Ig基因的序列特征与对不同流感毒株的抗体反应性联系起来。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接受者之间的Ig基因序列，以及Ig和Ig A亚型之间的序列。 相关性(见说明)：这项研究与流感疫苗的设计和管理直接相关，以预防大流行和流行性流感，特别是在幼儿和老年人中。它还将提供有关产生保护性B细胞免疫的机制的重要信息，以抵御广泛的其他重要人类病原体和生物恐怖主义因素。

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.