The B cell repertoire as a window into the nature and impact of the lung virome

项目摘要 呼吸道病毒对人类健康有重大影响。肺炎是住院的主要原因 这是非常年轻和老年人死亡的主要原因，最常见的是由呼吸道病毒引起的。 人类对由这些常见病原体引起的严重感染的易感性存在相当大的差异。 遇到了病毒，在机械水平上知之甚少。此外，肺炎的病因仍然是 这是一个谜，许多感染是由目前怀疑的微生物以外的不明微生物引起的。重复 暴露于呼吸道病毒对记忆淋巴细胞的数量有持久的影响， 在肺内，有助于控制这种局部动态病毒组。这包括记忆B细胞。的 通过克隆选择和亲和力成熟共同形成B细胞记忆， 病毒组相互作用的个体自然史。这种印记可以通过测序来检测。 肺B细胞免疫球蛋白可变区基因库及其克隆结构分析 统计学上。我们提出，鉴定被肺B细胞识别的具有最大亲和力成熟的病毒 将为病毒组对肺施加免疫压力提供一个公正的窗口。我们将研究 肺B细胞使用一套专门的方法，包括IgVRG库分析，一个创新的野岛 培养系统，用于从肺样本中培养B细胞克隆，以及新开发的多重VirScan 用于检测病毒特异性的技术，以鉴定产生最严重， 对人肺的复发性或慢性局部免疫压力。这些人类肺病毒组分析将 通过研究肺B细胞和呼吸道病毒如何因果相互作用来补充， 记忆B细胞在不同区室或条件下的差异，决定了局部免疫治疗的功效。 免疫控制肺部病毒。在R61阶段，我们将1）使用小鼠模型来检验假设 呼吸道病毒组动力学改造肺B细胞; 2）建立人Nojima B细胞培养体系 肺B细胞; 3）确定人肺是否含有可被VirScan检测到的病毒组反应性B细胞 技术.如果这些概念验证实验成功，那么在R33阶段，我们将测试 假设：4）由人肺病毒组和肺居民B组成的群体动力学系统 细胞与其他解剖部位不同; 5）人肺内存在异质性， 肺的中央、传导气道丰富的区域识别不同的病毒抗原， 6）人类肺病毒组与肺病毒之间的长期相互作用， 常驻B细胞有助于肺炎易感性; 7）肺记忆B细胞具有独特的 提供呼吸道病毒的快速局部抗体介导控制的特化表型。本研究 将利用肺驻留B细胞区室提供对肺病毒组及其 与肺免疫学和人类健康和疾病的关系。

Project summary Respiratory viruses have a major impact on human health. Pneumonia is the primary cause of hospitalizations of the very young and a leading cause of death in older adults, and is most often caused by respiratory viruses. There is considerable variation in the susceptibility of humans to severe infection caused by these commonly encountered viruses, poorly understood at mechanistic levels. Further, the etiology of pneumonia remains enigmatic, with many infections caused by unidentified microbes other than the current suspects. Repeated exposures to respiratory viruses have a lasting impact on populations of memory lymphocytes that reside within the lung, contributing to the control of this local dynamic virome. This includes memory B cells. The shaping of B-cell memory through clonal selection and affinity maturation together leaves a signature of the individual's natural history of virome interactions. This imprint can be detected by sequencing the immunoglobulin variable-region gene (IgVRG) repertoire of lung B cells and analyzing their clonal structure statistically. We propose that identifying viruses recognized by lung B cells with the greatest affinity maturation will provide an unbiased window into the virome exerting immunological pressure on the lung. We will study lung B cells using a suite of specialized approaches including IgVRG repertoire analyses, an innovative Nojima culture system for growing B cell clones out of lung samples, and the newly developed multiplex VirScan technology for detecting viral specificity, in order to identify the virome that has exerted the most severe, recurrent, or chronic local immunological pressure on the human lung. These human lung virome analyses will be complemented by studies of how lung B cells and respiratory viruses interact causally, to test whether there are differences in memory B cells across compartments or conditions that determine the efficacy of local immune control of viruses in the lung. In the R61 phase, we will 1) use a mouse model to test the hypothesis that respiratory virome dynamics remodel lung B cells; 2) establish the Nojima B cell culture system for human lung B cells; 3) determine if human lungs contain virome-reactive B cells that can be detected by VirScan technology. If these proof-of-concept experiments are successful, then in the R33 phase we will test the hypotheses: 4) that the population-dynamic system comprising the human lung virome and lung-resident B cells is distinctive from other anatomical sites; 5) that there is heterogeneity within the human lung, such that central, conducting airway-rich regions of the lung recognize different viral antigens than do those from more peripheral regions of the same lung; 6) that long-term interactions between the human lung virome and resident B cells contribute to pneumonia susceptibility; 7) that lung memory B cells have a unique and specialized phenotype that provides rapid local antibody-mediated control of respiratory viruses. This study will leverage the lung resident B-cell compartment to provide novel insights into the lung virome and its relationships to pulmonary immunology and human health and disease.