Vaccination and infection: indicators of immunological health and responsiveness

描述（由申请人提供）：在本提案中，我们的目标是使用系统生物学方法来调查一系列不同疫苗的免疫反应性，包括季节性流感和新的大流行性H1N1 v、带状疱疹、麻疹和脑膜炎奈瑟菌。在流感和带状疱疹的情况下，我们也将能够直接比较对疫苗的免疫反应与对病原体本身的自然反应。这些研究将比较不同的年龄组-儿童，年轻人和老年人-寻找特定的标记物和免疫能力的测定，特别是那些可能在不同的病原体和年龄组中常见的。我们还将调查免疫系统受损或失调的个体对流感疫苗的反应，以了解这些个体如何偏离健康个体。我们将采用一个简化的模型，其中暴露于不同疫苗或传染病的个体的临床标本将在我们的人类免疫监测中心进行一系列基本免疫功能的测定，并进行几种“最先进”的测定，例如我们最近开发的高通量HLA测序技术，该技术能够同时从数百人中生成完整的HLA单倍型。我们还将进行选定的免疫球蛋白和T细胞受体库分析和组合肽-MHC四聚体分析，以有效地搜索信息性T细胞表位。我们将采用先进的生物信息学分析，例如我们的新细胞类型特异性SAM算法（csSAM），该算法使用细胞亚群信息从全血基因表达数据中获得更高的灵敏度，并开发新的生物信息学工具来询问我们在所有项目，试点和核心中丰富而复杂的数据库。 相关性：总之，我们希望分析各种不同的疾病/疫苗模型，以定义应答者和非应答者个体的共同和独特特征，以开发新的信息工具和测定法，这些工具和测定法应该能够阐明关于这些研究组的特定问题，并且通常也将在免疫健康和功能障碍的“指标”方面受益。 项目1 标题：流感感染和疫苗接种中的浆母细胞运输和抗体应答 项目负责人：Greenberg，H 项目1描述（由申请方提供）：流感感染和疫苗接种中的浆母细胞运输和抗体应答。2009年的流感大流行再次强调了开发改良流感疫苗的紧迫性。这次大流行是由甲型H1N1流感变异株（H1N1 v）引起的，大部分人口对这种病毒几乎没有预先存在的免疫力。H1N1 v疫苗最近被批准在秋季分发。随着新的大流行性流感毒株的出现，这种非常罕见的情况提供了解决关于B细胞对流感的免疫力的几个关键问题的机会，这是保护免受流感感染的关键决定因素。针对流感感染或疫苗接种的B细胞应答的传统评价严重依赖于恢复期血清抗体测定，其可能不代表整个抗体应答，尤其是粘膜抗体应答，其通常对于保护免受呼吸道病原体非常重要。抗体应答首先由活化的B细胞或浆母细胞介导，其通过循环迁移到不同的靶位点并成为效应B细胞或浆细胞。最近，我们已经开发了一个全面的流式细胞术测定，以确定多种贩运受体的表达模式的blood plasmablasts，包括那些贩运信号的呼吸树。我们还开发了灵敏的方法来收集和分析由浆母细胞群体或其特定亚群分泌的多克隆抗体。利用这些新的检测方法，以及H1N1 v大流行提供的难得机会，我们将致力于以下具体目标：1.分析自然感染和两种流感疫苗诱导感染后浆母细胞上的转运受体谱; 2.定义不同年龄组中成浆细胞衍生的多克隆抗体（PPAb）对急性感染与粘膜或全身接种的应答的定量和定性差异; 3.定性和定量比较野生型H1N1 v感染诱导的同型和异型PPAb反应性与两种类型H1N1 v流感疫苗以及季节性流感疫苗诱导的PPAb反应性。这些目标将共同定义新的免疫指标，并为B细胞对流感感染和疫苗接种的反应机制提供新的见解。

DESCRIPTION (provided by applicant): In this proposal, we aim to use a systems biology approach to survey immune responsiveness across a range of different vaccines including influenza, both seasonal and the new pandemic H1N1v, Herpes zoster, Measles, and the bacterium Neisseria meningitidis. In the case of influenza and Herpes zoster, we will also be able to directly compare the immune response to vaccines with those natural responses to the pathogen itself. These investigations will compare different age groups-children, young adults, and the elderly-to look for specific markers and assays of immune competence and especially those that might be common across different pathogens and age groups. We will also survey the influenza vaccine responses of individuals with impaired or dysregulated immune systems in order to see how these might deviate from healthy individuals. We will employ a streamlined model in which clinical specimens from individuals exposed to different vaccines or infectious diseases will all be assayed for an array of basic immune functions in our Human Immune Monitoring Center and also for several "state of the art" assays such as our recently developed high throughput HLA sequencing technique, which is able to generate complete HLA haplotypes from hundreds of people simultaneously. We will also perform selected Immunoglobulin and T Cell Receptor repertoire analysis and combinatorial peptide-MHC tetramer analysis to efficiently search for informative T cell epitopes. We will employ advanced bioinformatics analysis such as our new cell type specific SAM algorithms (csSAM), which use cell subset information to achieve much higher sensitivity from whole blood gene expression data as well as developing new bioinformatics tools to interrogate our rich and complex data base across all projects, pilots and cores. RELEVANCE: In summary, we wish to analyze a variety of different disease/vaccine models in order to define common and unique characteristics of responder and non-responder individuals to develop new informative tools and assays that should be illuminating both to specific questions regarding these study groups and will also be of benefit generally with respect to "metrics" of immunological health and dysfunction. PROJECT 1 Title: Plasmablast Trafficking and Antibody Response in Influenza Infection and Vaccination Project Leader: Greenberg, H PROJECT 1 DESCRIPTION (provided by applicant): Plasmablast trafficking and antibody response in influenza infection and vaccination. The 2009 influenza pandemic reiterates the urgency in developing improved influenza vaccines. This pandemic is caused by the influenza A/H1N1 variant strain (H1N1v), to which much of the human population has little pre-existing immunity. H1N1v vaccines have recently been approved for distribution in the fall. This very uncommon circumstance with the advent of a new pandemic influenza strain provides the opportunity to address several critical questions regarding the B cell immunity against influenza, a key determinant of protection against influenza infection. Traditional evaluation for B cell responses against influenza infection or vaccination has relied heavily on convalescent serum antibody assays, which may not represent the entire antibody response, especially mucosal antibody responses that are frequently of great importance for protecting against respiratory pathogens. Antibody responses are first mediated by activated B cells, or plasmablasts, which migrate through circulation to different target sites and become effector B cells, or plasma cells. Recently we have developed a comprehensive flow cytometric assay to define the patterns of multiple trafficking receptor expression on blood plasmablasts, including those with trafficking signals for the respiratory tree. We have also developed sensitive methods to collect and analyze the polyclonal antibodies secreted by the plasmablast population or its specific subsets. Taking advantage of these new assays, and the rare opportunity provided by the H1N1v pandemic, we will address the following specific aims to: 1. Analyze the trafficking receptor profiles imprinted on plasmablasts after natural infection with those induced by immunization with two different influenza vaccines; 2. define the quantitative and qualitative differences in plasmablast-derived polyclonal antibody (PPAb) responses to acute infection vs. mucosal or systemic vaccination in different age groups; 3. qualitatively and quantitatively compare the homotypic and heterosubtypic PPAb reactivity induced by wild type H1N1v infection vs. the PPAb reactivity induced by the two types of H1N1v influenza vaccine, as well as by seasonal influenza vaccines. Together these aims will define new immune indicators and provide new insights to the mechanisms of B cell response to influenza infection and vaccination.