Multidimensional Single Cell Analysis of the Germinal Center Reaction

生发中心反应的多维单细胞分析

• 批准号: 9010478
• 负责人: Michael G. McHeyzer-Williams
• 金额: $ 60.23万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9010478
• 关键词: Adoptive Transfer; Affinity; Animals; Antibodies; Antibody Affinity; Antibody Specificity; Antigens; Area; B-Lymphocytes; Behavior; Binding; Biological Assay; Cell Count; Cell physiology; Cells; Clonal Expansion; Complex; Controlled Study; Cues; Evolution; Exposure to; Genetic Transcription; Goals; Immune; Immune system; Immunity; Immunoglobulin Class Switching; Infection; Life; Lymphoid Tissue; Measures; Mediating; Memory; Memory B-Lymphocyte; Methods; Molecular; Pathway interactions; Plasma Cells; Proteins; Reaction; Regulation; Repression; Research; Resolution; Scanning; Staging; Structure of germinal center of lymph node; Study models; Time; Transcriptional Regulation; Vaccination; Vaccine Antigen; Vaccine Design; Vaccine Research; Vaccines; Variant; adaptive immunity; base; design; human disease; immune function; immunogenicity; in vivo; lymph nodes; mouse model; pathogen; programs; public health relevance; response; single cell analysis; vaccine development

 DESCRIPTION (provided by applicant): Producing potent high-affinity antibodies that can neutralize target pathogens remains a central goal for vaccine research. While a great deal is known about antibody molecules themselves, still too little is understood about the programming of high-affinity memory B cells that produce them. Many promising vaccine antigens fail to elicit protective antibodies using contemporary vaccine formulations. Thus, major gaps remain in our basic understanding of antibody affinity maturation and how to enhance poor immunogenicity using rational protein vaccination. Research Focus: GCs are dynamic microenvironments that emerge within secondary lymphoid tissues following exposure to foreign antigens. The evolution of antibody specificity within these GC reactions is required to generate high-affinity memory B cells and long-lived plasma cells. Follicular helper T (TFH) cells provide the molecular cues for initiation of GCs and control of ongoing evolutionary mechanisms within the GC reaction. Understanding the regulatory mechanisms that control memory BCR evolution and GC B cell function remain an important basic goal for the field and is the major focus of the current application. Specific Aims: We have developed multi-dimensional single cell strategies to track these complex cellular behaviors at the clonal and sub-clonal levels in vivo. We will use this approach: SA-1: to study the molecular control of secondary GC induction and function at recall SA-2: to dissect the stage-specific cyclic progression of GC transcriptional programming SA-3: to modify the molecular control of GC B cell evolution and GC exit. Impact: We use polyclonal murine models of protein vaccination with state-of-the-art single cell analyses of antigen-specific immune function. These high-resolution studies of model antigens have the capacity to shift the basic conceptual framework that surrounds existing vaccine paradigms. Importantly, these basic new principles and assays can be used to re-design contemporary vaccine formulations that optimize high-affinity B cell immunity to more complex antigens.

 描述（由适用提供）：产生可以中和靶病原体的潜在高亲和力抗体仍然是疫苗研究的中心目标。尽管对抗体分子本身知之甚少，但关于产生它们的高亲和力记忆B细胞的编程仍然很少了解。许多有希望的疫苗抗原无法使用当代疫苗配方引起受保护的抗体。这是我们对抗体亲和力成熟的基本理解以及如何使用理性蛋白疫苗接种增强免疫原性的主要差距。研究重点：GC是动态的微环境，在暴露于外来抗原后次生淋巴组织内出现。这些GC反应中需要抗体特异性的演变才能产生高亲和力的记忆B细胞和长寿命的浆细胞。滤泡辅助辅助剂T（TFH）细胞提供了启动GC和控制GC反应中持续进化机制的分子线索。了解控制记忆BCR演化和GC B细胞功能的调节机制仍然是该领域的重要基本目标，并且是当前应用的主要重点。具体目的：我们已经开发了多维单细胞策略，以在体内在克隆和亚克隆水平上跟踪这些复杂的细胞行为。我们将使用这种方法：SA-1：研究次级GC诱导和回忆SA-2功能的分子控制：剖析GC转录编程SA-3的阶段特异性环状进程：修改GC B细胞演化和GC出口的分子控制。影响：我们使用抗原特异性的最先进的单细胞分析，使用蛋白质疫苗接种的多克隆鼠模型 免疫功能。这些对模型抗原的高分辨率研究具有改变现有疫苗范式的基本概念框架的能力。重要的是，这些基本的新原理和测定可用于重新设计当代疫苗配方，以优化对更复杂抗原的高亲和力B细胞免疫学。