Characterization of virus-specific human B cell subsets in lymphoid and non-lymphoid tissues

淋巴和非淋巴组织中病毒特异性人类 B 细胞亚群的表征

• 批准号: 10592418
• 负责人: Frances E. Lund
• 金额: $ 94.58万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10592418
• 关键词: Adipose tissue; Affinity; Alabama; Antibodies; Antibody titer measurement; B-Lymphocyte Subsets; B-Lymphocytes; Blood; Blood Cells; Bone Marrow; Cell physiology; Cellular biology; Clone Cells; Coupled; Data; Genetic Transcription; Goals; Helper-Inducer T-Lymphocyte; Hemagglutinin; Homing; Human; Immunity; Immunoglobulin Class Switching; Immunoglobulin-Secreting Cells; Infection; Influenza; Influenza Hemagglutinin; Knowledge; Ligands; Longevity; Lower respiratory tract structure; Lung; Lymphoid; Lymphoid Tissue; Memory B-Lymphocyte; Morbidity - disease rate; Mucous Membrane; Mus; Normal tissue morphology; Nucleoproteins; Organ; Organ Donor; Phenotype; Production; Property; Proteins; Publishing; Recombinants; Sialic Acids; Site; Sorting; Specificity; Spleen; Structure of parenchyma of lung; System; Testing; Tissues; Upper respiratory tract; Vaccination; Vaccine Design; Viral; Virus; Visceral; acute infection; cross reactivity; cytokine; design; experimental study; human tissue; influenza virus vaccine; influenzavirus; innovation; lymph nodes; lymphoid organ; mortality; peripheral blood; programs; response; viral transmission

Project 2: Characterization of Virus-Specific Human B Cell Subsets in Lymphoid and Non-Lymphoid Tissues Project Summary Antibodies against influenza virus protect against infection and reduce morbidity and mortality. As a result, vaccines against influenza are designed to elicit antibodies against circulating strains of virus. Given that antibodies are made by B cells, it makes sense to characterize influenza-specific B cells in order to determine how they are selected, how they function and where they are maintained. However, most studies of influenza- specific B cells in humans are limited to B cells from peripheral blood, rather than those in lymphoid organs or the lung. Moreover, most studies characterize influenza-specific antibody-secreting cells (ASCs), which circulate for a short period after infection or vaccination. Consequently, we have minimal understanding of influenza-specific memory B cells or ASCs in human tissues. Thus, there is an urgent need to characterize the phenotypes and functions of influenza-specific B cells in human lymphoid and non-lymphoid tissues. Given this gap in knowledge, we are proposing experiments that will use “B cell tetramers” to identify influenza-specific B cells responding to hemagglutinin (HA), nucleoprotein (NP) and non-structural-1 (NS1) proteins of influenza virus in blood, lymphoid tissues, lung and visceral adipose tissues of normal human donors. We will then determine how the phenotypes, transcriptional programs, functional properties, reactivities and affinities of those B cells differ between tissues and between B cells of different specificities. Importantly, we will use an automated, single-cell cloning and antibody-expression system to rapidly and efficiently generate recombinant, influenza-specific antibodies from sorted memory B cells in order to determine their affinity and cross-reactivity. Therefore, we believe that this project is highly innovative and will significantly advance our understanding of influenza-specific B cell biology in various human tissues.

项目2：类淋巴细胞和非类淋巴细胞中病毒特异性人B细胞亚群的表征 组织 项目摘要 抗流感病毒的抗体可防止感染并降低发病率和死亡率。因此，在本发明的一个方面， 针对流感的疫苗被设计为引发针对病毒的循环株的抗体。鉴于 抗体是由B细胞产生的，因此有必要对流感特异性B细胞进行表征，以确定 它们是如何被选择的，它们是如何运作的，以及它们在哪里被维护。然而，大多数流感研究- 人的特异性B细胞限于来自外周血的B细胞，而不是淋巴器官或 肺此外，大多数研究描述了流感特异性抗体分泌细胞（ASC）， 在感染或接种疫苗后短时间内传播。因此，我们对 流感特异性记忆B细胞或人组织中的ASC。因此，迫切需要确定 人淋巴和非淋巴组织中流感特异性B细胞的表型和功能。鉴于这种 由于知识上的差距，我们建议使用“B细胞四聚体”来鉴定流感特异性B 对流感病毒的血凝素（HA）、核蛋白（NP）和非结构蛋白1（NS 1）蛋白应答的细胞 正常人供体的血液、淋巴组织、肺和内脏脂肪组织中的病毒。然后我们将 确定表型，转录程序，功能特性，反应性和亲和力， 这些B细胞在组织之间和在不同特异性的B细胞之间是不同的。重要的是，我们将使用 自动化的单细胞克隆和抗体表达系统，以快速有效地产生重组体， 从分选的记忆B细胞中分离流感特异性抗体，以确定它们的亲和力和交叉反应性。 因此，我们认为，该项目具有很强的创新性，将大大促进我们对 流感特异性B细胞生物学。