Visualizing the signals that drive resident memory T cell formation in skin

项目摘要 皮肤不仅是一个物理屏障，而且还窝藏各种先天性和适应性免疫细胞 这些类型共同维持功能性屏障以抵抗肠道微生物的入侵， 环境病原体这一屏障功能必须与避免夸大或 不适当的免疫反应可诱发皮肤免疫病理学。 CD 8+表皮驻留记忆T细胞（eTrm）最近被认为是免疫调节的关键组成部分。 皮肤免疫防御然而，除了保护我们免受以前遇到的再次感染外， 病原体，它们也可能在几种自身免疫性和炎症性皮肤中发挥重要的致病作用。 疾病，如牛皮癣、接触性皮炎、慢性湿疹等。了解eTrm如何 因此，形成和维持的皮肤炎症与皮肤病的新治疗策略的发展有关。 疾病 遗传小鼠模型在鉴定关键蛋白质和分子机制方面具有高度指导性， 在包括皮肤在内的各种上皮组织中介导eTrm分化。然而，他们还没有生产 描述连续迁移步骤的综合模型，通过该模型，在炎症反应期间， 一些效应T细胞被保留在皮肤中，被引导穿过上皮基底膜进入皮肤。 表皮，并进入壁龛，促进长期局部持久性作为局部记忆的能力 细胞这阻碍了对这一过程的全面理解，而这是理性思考所必需的。 设计治疗干预措施，以增强疫苗接种背景下的eTrm形成或破坏 它们在皮肤免疫病理学背景下的形成。 我们建议开发一种多光子活体显微镜方法， 表征迁移事件，而不是促进小鼠皮肤中的eTrm分化过程。我们将评估 对细胞因子TGF-β不敏感的CD 8 T细胞的迁移行为如何与野生型不同 细胞，并使用荧光融合蛋白报告构建体，以真实的时间可视化T细胞如何瞬时 收集TGF-β信号，从而探索这些关键信号事件发生的背景。最后， 我们将通过测试TGF-β调节基因CD 103和S1 P-receptor-1的功能来确定它们的功能。 在TGF-β不敏感的T细胞中恢复eTrm分化方面的能力。

Project Summary The skin not only serves as a physical barrier, but also harbors a variety of innate and adaptive immune cell types that collectively maintain a functional barrier against the invasion of commensal microbes and environmental pathogens. This barrier function has to be balanced with the need to avoid exaggerated or inappropriate immune responses that can induced skin immunopathology. CD8+ epidermal resident memory T cells (eTrm) have recently become recognized as a critical component of skin immune defense. However, in addition to protecting us against re-infection with previously encountered pathogens, they likely also play important pathogenic roles in several autoimmune and inflammatory skin diseases, such as psoriasis, contact dermatitis, chronic eczema, and others. Understanding how eTrm are formed and maintained is therefore relevant to the development of novel therapeutic strategies for skin disease. Genetic mouse models have been highly instructive in identifying key proteins and molecular mechanisms that mediate eTrm differentiation in various epithelial tissues, including skin. However, they have not yet produced a comprehensive model describing the sequential migratory steps, by which during an inflammatory response some effector T cells in are retained in skin, guided to cross the epithelial basement membrane to enter the epidermis, and settle into niches that promote the capacity for long-term local persistence as local memory cells. This has hindered an integrated understanding of this process, which will be required for the rational design of therapeutic interventions to either enhance eTrm formation in the context of vaccination or disrupt their formation in the context of skin immunopathology. He we propose to develop a multiphoton intravital microscopy approach to visualize and comprehensively characterize migratory events than facilitate the process of eTrm differentiation in mouse skin. We will assess how CD8 T cells that are insensitive to the cytokine TGF-beta diverge in their migratory behavior from wildtype cells, and use fluorescent fusion protein reporter constructs to visualize in real time how T cell instantaneously collect TGF-beta signals and thereby explore the context in which these critical signaling events occur. Finally, we will determine the function of the TGF-beta-regulated genes CD103 and S1P-receptor-1 by testing their ability to restore aspects of eTrm differentiation in TGF-beta-insensitive T cells.