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，从而 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 多回螺旋体寄生虫(HP)，是公认的2型免疫诱导剂。在这 系统中，小鼠对初次感染治疗后的再感染具有抵抗力，抵抗力为Th2。 依赖细胞。我们最近报道，在Hp感染期间，肠系膜脂肪组织(MAT)变成 由长寿命的Th2rM细胞填充，这些细胞除了签名的Th2外还产生双调蛋白和转化生长因子β1 细胞因子IL-4、IL-5和IL-13以及基质细胞上两性调节蛋白受体的缺失与 对Hp的易感性增加。此外，垫内的间质细胞，特别是具有 多能潜能(多能祖细胞，MPC)分化为成纤维细胞或脂肪细胞，成为 激活并使警报因子IL-33和TSLP能够促进Th2RM细胞的激活。我们有 Th2RM细胞和MPC均定位于MAT的间质间隙。我们建议MAT与 肠道参与对幽门螺杆菌的保护性反应。这涉及到生产警报器以 支持Th2RM细胞的活动，并产生细胞外基质(ECM)，我们假设这对 在肉芽肿中捕获入侵的寄生虫，并在感染期间支持软组织完整性和修复。 根据我们已发表的发现和新的初步数据，我们假设Th2RM细胞和基质细胞 在HP期间，团队合作，相互促进对方的激活，发挥宿主保护作用 感染。根据我们最近发表的关于MPC先天训练的发现和新的初步数据， 对于Th2RM细胞的脂代谢和运动，我们制定了两个特定的目标：1，了解 MPC在形成宿主对Hp的抵抗力中的作用；2，确定其功能、抗原特异性和细胞- MAT驻留的Th2细胞的内在代谢和迁移特征。为了解决这些问题，我们将使用RNA SEQ、ATACseq、流式细胞术、寄生虫学技术、细胞体外运动的高级成像 组织硬度的测量，Th2RM细胞和基质细胞功能特性的体外研究， 体内功能丧失模型，探讨双调蛋白、转化生长因子β、IL-33和TSLP的作用。我们的工作已经完成 揭示基质细胞生物学的新特征的潜力，这些特征是潜在的效应机制所不可或缺的 对肠道病原体的抵抗力和免疫力，并强调潜在的干预要点 操纵基质细胞生物学，因为它与健康和疾病中的2型免疫有关。我们的发现可能有 与了解人类状况的相关性，如过敏/过敏、伤口愈合受损、纤维化和 癌症，与基质细胞生物学失调和细胞外基质的产生有关。

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.