Duration and Function of Lung Tissue Resident CD8+ Memory T cells

项目总结/摘要 流感是一种呼吸道病毒，估计每年感染全球10亿人。虽然 是一种流感疫苗，通常只有40-60%的有效性，人们必须每年接种。此外，本发明还 该疫苗不能预防未来的流感，可能是大流行性流感，因此有一个主要的 需要新的疫苗接种策略。CD 8 + T细胞可以提供针对不同菌株的交叉保护， 因为它们识别保守的内部表位。具体而言，由于其在 肺，CD 8+组织驻留记忆T细胞（TRM）可以迅速响应感染和介导的保护， 降低病毒载量和免疫病理学。在小鼠中，研究表明流感特异性CD 8+肺TRM TRM的耐久性有限，但尚不清楚肺TRM在人类中存活多久。此外， 哪些肺TRM细胞因子降低小鼠的病毒载量和免疫病理学尚不清楚， 关于人类肺TRM的功能。在这里，我们提出了一系列的实验，以填补这些关键的差距， 知识在目标1中，我们将使用来自肺移植患者的纵向支气管肺泡灌洗样本， 特异于受体和供体之间不匹配的HLA等位基因的抗体，以跟踪供体肺移植的持续时间， TRM和受体肺TRM的发展，通过流式细胞术。此外，我们将确定动态 通过对纵向支气管肺泡灌洗，血液， 以及来自感染流感的肺移植患者的鼻灌洗样本。在目标2中，我们将 获得不能移植的健康肺，以检查人肺的效应器功能 TRM。将用流感肽库刺激肺细胞，以确定流感病毒产生的细胞因子。 特异性肺TRM，以及这些细胞因子对肺组织中邻近细胞的影响。另外我们 然后将使用小鼠来机械地确定单个肺TRM细胞因子降低病毒载量的效果 激活邻近的先天细胞通过这项拟议的工作，我们希望更好地了解 人肺TRM在对流感的免疫应答中起作用。最后，由于肺TRM是响应于 呼吸道病毒，而不仅仅是流感，这项研究的结果可以提供对免疫系统的见解 这可以改进针对所有呼吸道病毒的疫苗设计，并有助于预防未来的全球流行病。

PROJECT SUMMARY/ABSTRACT Influenza is a respiratory virus that infects an estimated one billion people around the world annually. While there is an influenza vaccine, it is typically only 40-60% effective and people must receive it every year. Additionally, the vaccine does not protect against future, and possibly pandemic, strains of influenza and thus there is a major need for new vaccination strategies. CD8+ T cells can provide cross protection against different strains of influenza because they recognize internal epitopes that are conserved. Specifically, due to their position in the lung, CD8+ tissue resident memory T cells (TRM) can rapidly respond to infection and mediate protection by reducing viral loads and immunopathology. In mice, studies have shown that influenza-specific CD8+ lung TRM have limited durability, but it is unknown how long lung TRM survive in humans. Additionally, the mechanisms by which lung TRM cytokines reduce viral loads and immunopathology is unknown in mice, and even less is known about the function of lung TRM in humans. Here, we propose a series of experiments to fill these critical gaps in knowledge. In Aim 1, we will use longitudinal bronchoalveolar lavage samples from lung transplant patients and antibodies specific for mismatched HLA alleles between recipient and donor to track the duration of donor lung TRM and the development of recipient lung TRM by flow cytometry. Additionally, we will determine the dynamics of flu-specific human lung TRM by performing immunophenotyping on longitudinal bronchoalveolar lavage, blood, and nasal lavage samples from lung transplant patients that become infected with influenza. In Aim 2, we will obtain healthy lungs that are unable to be transplanted in order to examine the effector functions of human lung TRM. Lung cells will be stimulated with influenza peptide pools to determine the cytokines produced by influenza- specific lung TRM, and the impact those cytokines have on neighboring cells in the lung tissue. Additionally, we will then use mice to mechanistically determine the effect of individual lung TRM cytokines to reduce viral loads and activate neighboring innate cells. Through this proposed work we hope to better understand the role that human lung TRM play in the immune response to influenza. Lastly, as lung TRM are generated in response to respiratory viruses, and not just influenza, results of this study could provide insights into the immune system that could improve vaccine design for all respiratory viruses and aid in the prevention of future global pandemics.