Stromal cells in immunity to infection

项目摘要 我们工作的目标是破译基质细胞如何与免疫细胞相互作用，以改变组织生物学， 支持抵抗肠道感染。我们的工作将使用感染肠道蠕虫的小鼠 寄生虫Heligmosomoides polygyrus bakeri（Hp），其是公认的2型免疫诱导剂。在这 系统中，小鼠对原发感染的治愈性治疗后的再感染具有抗性，并且抗性是Th 2 依赖细胞的我们最近报道，在Hp感染期间，肠系膜脂肪组织（mAT）变得 由长寿的Th 2 RM细胞组成，除了Th 2细胞外，还产生双调蛋白和TGFβ1 细胞因子IL-4、IL-5和IL-13，基质细胞上双调蛋白受体的缺失与 增加对Hp的易感性。此外，mAT内的基质细胞，特别是具有mAT的基质细胞亚群， 多能潜能（多能祖细胞，MPC）分化成成纤维细胞或脂肪细胞，成为 活化并产生能够促进Th 2 RM细胞活化的alarmins IL-33和TSLP。我们有 在mAT中，Th 2 RM细胞和MPC均定位于间质空间。我们认为mAT与 肠参与对Hp的保护性反应。这涉及到生产警报， 支持Th 2 RM细胞活性，并产生细胞外基质（ECM），我们推测这对于 在肉芽肿中捕获入侵的寄生虫，并在感染期间支持软组织的完整性和修复。 基于我们已发表的研究结果和新的初步数据，我们假设Th 2 RM细胞和基质细胞 作为一个团队，在Hp期间相互促进彼此的激活并发挥宿主保护作用 感染根据我们最近发表的关于MPC先天训练的研究结果和新的初步数据， 和脂质代谢以及Th 2 RM细胞的运动性，我们开发了两个具体的目的：1，了解 MPC在形成宿主对Hp的抗性中的作用; 2.确定MPC的功能、抗原特异性和细胞- mAT驻留的Th 2细胞的内在代谢和迁移特征。为了实现这些目标，我们将使用RNA seq、ATACseq、流式细胞术、寄生虫学技术、离体细胞运动的高级成像， 组织硬度的测量，Th 2 RM细胞和基质细胞的功能特性的离体研究， 以及体内功能丧失模型以探索双调蛋白、TGFβ、IL-33和TSLP的作用。我们的工作 揭示基质细胞生物学的新特征的潜力是潜在效应机制的组成部分 对肠道病原体的抵抗力和免疫力，并强调潜在的干预点， 操纵基质细胞生物学，因为它涉及健康和疾病中的2型免疫。我们的发现可能 与理解人类疾病的相关性，如特应性/过敏、伤口愈合受损、纤维化和 癌症，其中涉及失调的基质细胞生物学和ECM产生。

Project Summary The objective of our work is to decipher how stromal cells interact with immune cells to alter tissue biology to support resistance to an intestinal infection. Our work will use mice infected with the intestinal helminth parasite Heligmosomoides polygyrus bakeri (Hp), which is a recognized inducer of type 2 immunity. In this system, mice are resistant to reinfection following curative treatment of primary infection, and resistance is Th2 cell-dependent. We recently reported that during Hp infection mesenteric adipose tissue (mAT) becomes populated by long-lived Th2RM cells which make Amphiregulin and TGFβ1 in addition to the signature Th2 cytokines IL-4, IL-5 and IL-13, and that deletion of the Amphiregulin receptor on stromal cells is associated with increased susceptibility to Hp. Further, stromal cells within mAT, and especially a subset of stromal cells with multipotent potential (multipotent progenitor cells, MPC) to differentiate into fibroblasts or adipocytes, become activated and makes the alarmins IL-33 and TSLP that are able to promote Th2 RM cell activation. We have localized both Th2RM cells and MPC to interstitial spaces in mAT. We propose that mAT interacts with the intestine to participate in the protective response against Hp. This involves the production of alarmins to support Th2RM cell activity, and to produce extracellular matrix (ECM), which we postulate is important for trapping invading parasites in granulomas, and supporting soft tissue integrity and repair during infection. Based on our published findings and new preliminary data, we hypothesize that Th2RM cells and stromal cells work as a team to mutually facilitate each other’s activation and exert host protective effects during Hp infection. On the basis of our recently published findings and new preliminary data on innate training in MPC, and lipid metabolism and motility of Th2RM cells, we have developed two Specific Aims: 1, To understand the role of MPC in shaping host resistance to Hp; and 2, To determine the function, antigen specificity, and cell- intrinsic metabolic and migratory features of mAT-resident Th2 cells. To address these Aims we will use RNA seq, ATACseq, flow cytometry, parasitological techniques, advanced imaging of cell movement ex-vivo, measurements of tissue stiffness, ex-vivo studies of the functional properties of Th2RM cells and stromal cells, and in vivo loss of function models to probe the roles of Amphiregulin, TGFβ, IL-33 and TSLP. Our work has the potential to reveal novel features of stromal cell biology that are integral to underlying effector mechanisms of resistance and immunity to intestinal pathogens, and to highlight potential points of intervention for manipulating stromal cell biology as it relates to type 2 immunity in health and disease. Our findings may have relevance to understanding human conditions such as atopy/allergy, impaired wound healing, fibrosis and cancer, in which dysregulated stromal cell biology and ECM production are implicated.