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

项目2：淋巴和非淋巴中病毒特异性人类B细胞亚群的特征 组织 项目摘要 针对流感病毒的抗体可预防感染，降低发病率和死亡率。结果, 流感疫苗的设计是为了激发针对流行病毒株的抗体。考虑到 抗体是由B细胞制造的，因此有必要对流感特异性B细胞进行鉴定，以确定 它们是如何选择的，它们如何发挥作用，以及在哪里维护它们。然而，大多数关于流感的研究- 人类的特异性B细胞仅限于外周血中的B细胞，而不是淋巴器官或 肺部。此外，大多数研究的特征是流感特异性抗体分泌细胞(ASCs)，它 在感染或接种疫苗后短时间内循环。因此，我们对此只有很少的了解 人类组织中的流感特异性记忆B细胞或ASCs。因此，迫切需要确定 人淋巴组织和非淋巴组织中流感特异性B细胞的表型和功能。鉴于此， 由于知识上的差距，我们提议进行实验，利用“B细胞四聚体”来识别流感特异的B细胞 对流感血凝素(HA)、核蛋白(NP)和非结构蛋白1(NS1)蛋白反应的细胞 正常人血液、淋巴组织、肺和内脏脂肪组织中的病毒。到时候我们会的 确定表型、转录程序、功能特性、反应性和亲和力 这些B细胞在不同组织和不同特性的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，快速高效地产生重组， 从分离的记忆B细胞中提取流感特异性抗体，以确定它们的亲和力和交叉反应性。 因此，我们相信这个项目具有很高的创新性，并将极大地促进我们对 人类不同组织中的流感特异性B细胞生物学。

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.