Unraveling Human T Follicular Helper Cell Development

揭开人类滤泡辅助 T 细胞发育的谜团

基本信息

批准号：
10568500
负责人：
Philip A Mudd
金额：
$ 58.12万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-12-06 至 2027-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10568500
关键词：
2019-nCoV Activities of Daily Living Address Adoptive Transfer Affinity Animal Model Antibodies Antibody Affinity Antigens Autologous Axillary lymph node group B-Cell Development B-Lymphocytes Biological Assay Biological Models Blood CD4 Positive T Lymphocytes COVID-19 COVID-19 pandemic COVID-19 vaccine Cell Maturation Cell Separation Cell physiology Clinical Collaborations Communicable Diseases Data Data Set Development Epitopes Event Exhibits Fine needle aspiration biopsy Flow Cytometry Genetic Transcription Helper-Inducer T-Lymphocyte Human Human Volunteers Immune response Immunoglobulin Class Switching Immunologist Infection Influenza vaccination Intramuscular Knowledge Lead Maintenance Measures Memory Memory B-Lymphocyte Methods Modeling Mus Participant Phenotype Plasma Cells Play Population Process Production Productivity Proteins RNA vaccination RNA vaccine Reaction Role SARS-CoV-2 spike protein Sampling Series Signal Transduction Site Sorting Specificity Structure of germinal center of lymph node Supporting Cell Surface System T cell receptor repertoire sequencing T-Cell Receptor T-Lymphocyte Subsets Techniques Testing Time Tissues Tonsillectomy Utah Vaccination Vaccines Variant Work antigen challenge cohort cytokine deltoid muscle draining lymph node gene function high dimensionality human model in vivo lipid nanoparticle lymph nodes mouse model neutralizing antibody novel plasma cell development recruit response seasonal influenza secondary lymphoid organ single-cell RNA sequencing time interval ultrasound vaccine distribution vaccine platform

2019-nCoV Activities of Daily Living Address Adoptive Transfer Affinity Animal Model Antibodies Antibody Affinity Antigens Autologous Axillary lymph node group B-Cell Development B-Lymphocytes Biological Assay Biological Models Blood CD4 Positive T Lymphocytes COVID-19 COVID-19 pandemic COVID-19 vaccine Cell Maturation Cell Separation Cell physiology Clinical Collaborations Communicable Diseases Data Data Set Development Epitopes Event Exhibits Fine needle aspiration biopsy Flow Cytometry Genetic Transcription Helper-Inducer T-Lymphocyte Human Human Volunteers Immune response Immunoglobulin Class Switching Immunologist Infection Influenza vaccination Intramuscular Knowledge Lead Maintenance Measures Memory Memory B-Lymphocyte Methods Modeling Mus Participant Phenotype Plasma Cells Play Population Process Production Productivity Proteins RNA vaccination RNA vaccine Reaction Role SARS-CoV-2 spike protein Sampling Series Signal Transduction Site Sorting Specificity Structure of germinal center of lymph node Supporting Cell Surface System T cell receptor repertoire sequencing T-Cell Receptor T-Lymphocyte Subsets Techniques Testing Time Tissues Tonsillectomy Utah Vaccination Vaccines Variant Work antigen challenge cohort cytokine deltoid muscle draining lymph node gene function high dimensionality human model in vivo lipid nanoparticle lymph nodes mouse model neutralizing antibody novel plasma cell development recruit response seasonal influenza secondary lymphoid organ single-cell RNA sequencing time interval ultrasound vaccine distribution vaccine platform

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项目摘要

ABSTRACT Neutralizing antibodies are critical for protection from infectious diseases. The lymph node (LN) germinal center (GC) is the site where B cells undergo antibody affinity maturation and develop into long-lived plasma cells – key events that are required for the development of highly effective neutralizing antibodies following infection or vaccination. T follicular helper cells (TFH) are the CD4+ T cell subset responsible for providing B cell help during an ongoing GC. TFH are absolutely required for GC formation and maintenance. By extension, TFH are necessary to produce effective neutralizing antibodies following antigen exposure. While many aspects of the TFH response and TFH function have been examined in animal models, human TFH responses in the draining LN have only recently been explored with the novel application of an established technique: serial ultrasound-guided fine needle aspiration of draining LN following vaccination. With this technique, we recently demonstrated that the antigen-specific TFH response to SARS-CoV-2 spike (S) protein mRNA vaccination persists in the GC for more than 4 months following vaccination and correlates with the presence of S-specific GC B cells. Furthermore, we have noted substantial LN TFH transcriptional phenotypic changes suggestive of functional maturation over this prolonged GC time interval using single cell RNA-seq in a small preliminary cohort. In this proposal, we will expand upon these findings to address our primary hypothesis: human TFH phenotypic maturation occurs over time in the draining LN following vaccination and these phenotypic changes are associated with changes in TFH function. To explore this hypothesis, we propose three specific aims: 1) We will first establish that TFH phenotypic maturation occurs over time in multiple antigen-specific TFH populations that we will define and characterize from a cohort of fourteen COVID-19 mRNA vaccine recipients using single cell RNA-seq and ex vivo epitope identification methods. 2) We will determine how these antigen-specific TFH populations change in a tertiary immune response following COVID-19 mRNA vaccine “boost” and continued serial LN sampling of the same cohort participants. We will also ascertain if new antigen-specific TFH populations are recruited to the GC during a recall response. 3) Finally, we will verify that the profound phenotypic changes we observe lead to changes in the functional capacity of antigen-specific TFH to provide help to B cells using an ex vivo system of sorted human LN TFH and an in vivo murine adoptive transfer model. By addressing these aims, we will significantly enhance our understanding of the role that human TFH play in directing GC B cell responses to vaccination.

抽象的中和抗体对于免受传染病的保护至关重要。淋巴结（LN）生发中心（GC）是B细胞经历抗体亲和力成熟并发展为长寿命的浆细胞的部位 - 关键在感染后开发高效中和抗体所需的事件或疫苗接种。 T卵泡辅助细胞（TFH）是CD4+ T细胞子集，负责在期间提供B细胞帮助持续的GC。 TFH是GC组建和维护绝对必需的。通过扩展，TFH是必要的在抗原暴露后产生有效的中和抗体。而TFH响应的许多方面在动物模型中已经检查了TFH功能，在排水LN中的人类TFH反应只有最近，通过新的应用技术的新应用探索了：串行超声引导罚款疫苗接种后排干LN的针头抽吸。通过这种技术，我们最近证明了抗原特异性TFH对SARS-COV-2尖峰（S）蛋白mRNA疫苗接种的反应持续在GC中以获取更多疫苗后4个月比S特异性GC B细胞的存在相关。此外，我们已经注意到大量的LN TFH转录表型变化暗示了功能成熟在小的初步队列中使用单细胞RNA-Seq长时间的GC时间间隔。在此提案中，我们将扩展这些发现以解决我们的主要假设：人类TFH表型成熟发生于疫苗和这些表型变化后排水LN的时间与TFH的变化有关功能。为了探讨这一假设，我们提出了三个具体目标：1）我们首先确定TFH表型随着时间的流逝，成熟发生在多个抗原特异性的TFH种群中使用单细胞RNA-Seq和Ex Vivo表位组成的14个Covid-19 mRNA疫苗受体识别方法。 2）我们将确定这些抗原特异性TFH种群如何变化 COVID-19 MRNA疫苗后的免疫反应“增强”和持续的串行LN采样队列参与者。我们还将确定在期间是否将新的抗原特异性TFH种群招募到GC 召回回应。 3）最后，我们将验证我们观察到的深刻的表型变化导致变化抗原特异性TFH的功能能力使用分类的人体系统为B细胞提供帮助 LN TFH和体内鼠自适应转移模型。通过解决这些目标，我们将大大增强我们对人类TFH在指导GC B细胞对疫苗接种反应中的作用的理解。