Granulysin and the antimicrobial activity of CD8T cells - development of a better model

抽象的。CD8 T细胞是获得性免疫的重要手臂，对 针对病毒、其他细胞内病原体和肿瘤的保护。我们的实验室有一个历史悠久的 对CD8 T细胞在结核病免疫中的作用感兴趣，结核病是导致死亡的主要原因 是世界上最大的传染病。来自非人类灵长类(NHP)结核病模型的最新数据 表明CD8T细胞在初级免疫和疫苗介导的免疫中都起着至关重要的作用 抗结核分枝杆菌[1-5]。CD8 T细胞数量和功能的增加是 与NHP结核病模型中疫苗诱导的保护有关。此外，使用单个电池 RNAseq、细胞毒性NK和CD8T细胞与宿主受益的结果显著相关(即， MTB控制)。重要的是，CTL经常与限制性肉芽肿有关。 表达穿孔素、颗粒酶和颗粒溶素的组合。颗粒溶素(GNLY)是一种重要的 由细胞毒性CD8 T细胞和NK细胞产生的细胞毒性颗粒成分，可杀死许多人 不同类型的细胞内细菌病原体。颗粒溶素蛋白具有抗菌活性 当直接应用于细菌时，可以对抗结核杆菌，并可以杀死耐药的结核杆菌菌株[6]。然而，它 一直很难研究颗粒溶素在小鼠模型中的作用，因为啮齿动物缺乏颗粒溶素的同源基因 基因。Allan Krensky[7]培育了一种人颗粒溶素转基因小鼠(hGNLY-TG)。在……里面 小鼠颗粒溶素主要表达于NK细胞，而CD8T细胞仅在旺盛生长后才表达。 体外刺激。在这里，我们建议建立新的小鼠模型来研究颗粒溶素在 抗菌免疫。我们的方法将解决颗粒溶素的表达是否对CD8T至关重要 结核分枝杆菌的细胞控制。我们的表达系统将能够更好地模拟人类CD8 T细胞 小鼠的反应。该模型将提供一个支持研究的实验系统 颗粒溶素在结核病免疫中的作用。我们假设，次优的抗菌功能 CD8T细胞在小鼠模型中缺乏颗粒溶素的表达。推论就是颗粒溶素 小剂量小鼠CD8T细胞表达可提高其控制结核分枝杆菌感染的能力 气溶胶结核分枝杆菌感染。作为这个项目的一部分，我们将回答这个问题。

Abstract. CD8 T cells are an important arm of adaptive immunity, and make critical contributions to protection against viruses, other intracellular pathogens, and tumors. Our lab has a long-standing interest in the role of CD8 T cells in immunity to tuberculosis (TB), which is the leading cause of death in the world from an infectious agent. Recent data from the non-human primate (NHP) model of TB shows that CD8 T cells make a crucial contribution to both primary and vaccine-mediated immunity against Mycobacterium tuberculosis (Mtb) [1-5]. Increases in CD8 T cell number and function are associated with vaccine-induced protection in the NHP TB model. Furthermore, using single cell RNASeq, cytotoxic NK and CD8 T cells are significantly associated with host-beneficial outcome (i.e., Mtb control). Importantly, CTL frequently associated with restrictive granulomas more frequently express the combination of perforin, granzymes, and granulysin. Granulysin (GNLY) is an important cytotoxic granule component produced by cytotoxic CD8 T cells and NK cells, and can kill many different types of intracellular bacterial pathogens. The granulysin protein has antimicrobial activity against Mtb when directly applied to bacteria and can kill drug resistant Mtb strains [6]. However, it has been difficult to study the role of granulysin in the mouse model, as rodents lack an ortholog of the gene. A human granulysin transgenic mouse (hGNLY-tg) was developed by Allan Krensky [7]. In that mouse, granulysin is expressed primarily in NK cells, and in CD8 T cells only after vigorous in vitro stimulation. Here, we propose to develop new mouse models to study the role of granulysin in antimicrobial immunity. Our approach will address whether granulysin expression is crucial for CD8 T cell control of Mtb. Our expression system will enable better modelling of human CD8 T cell responses in the mouse. This model will provide an experimental system to support investigation of the role of granulysin in immunity to TB. We hypothesize that the suboptimal antibacterial function of CD8 T cells in the murine model is their lack of granulysin expression. A corollary is that granulysin expression by murine CD8 T cells would improve their ability to control Mtb infection after low dose aerosol Mtb infection. We will answer this question as part of this project.