Cell-cell interactions driving gut inflammation and tolerance

项目摘要 高度特化的肠道免疫系统负责维持对来自肠道的无害刺激的耐受性。 食物和肠道细菌，同时提供对病原体的保护性免疫。这种失调 临界平衡可导致食物过敏、炎症性肠病（IBD）或增加对肠道疾病的易感性。 病原体树突状细胞（DC）是肠道内稳态的关键参与者， 通过呈递管腔抗原并诱导CD 4 + T细胞功能性分化为调节性T细胞， 或促炎亚群。决定耐受或免疫的细胞机制 肠抗原的免疫应答仍然未知，主要是因为确切的DC的鉴定和表征 在这些过程中所涉及的是一个长达数十年的技术挑战。为了解决这个问题，我 LIPSTIC（Labeling Immune Partnerships by SorTagging Intercellular Contacts） 该技术在肠道中，这使得特定的肠道细胞-细胞相互作用的邻近依赖性标记。的 这项研究的主要目标是阐明决定DCs是否诱导 调节性或促炎性T细胞，通过将LIPSTIC系统与基因靶向、功能性 成像、交叉遗传学和基于相互作用的转录组学方法。该提案测试了 假设DCs在肠道中存在不同的功能， 通过管腔内容物对DC亚群的重编程决定了对食物和微生物的耐受性或免疫性。 具体来说，这一概念将在生理（对食物和微生物的耐受性）和 病理（过敏性致敏、肠道感染和结肠炎）情况。为此，K99指导阶段 (Aim 1）将揭示诱导对食物的耐受性或炎症的DC的确切性质，以及细胞和 食物特异性PTB维持口服耐受性的分子机制。接下来，R 00独立阶段 (Aim 2）将专注于研究免疫反应的药物，与相关的疾病，如结肠炎。 阐明DC群体、位置和参与对不同微生物耐受的分子机制 将揭示健康和疾病中宿主-微生物相互作用的本质。这项研究计划将 为某些肠道疾病的病因学提供了基本的见解，并将为 食物过敏、结肠炎和肠道感染的新疗法的开发，对公众具有深远的意义。 健康拟议的发展计划补充了我在粘膜免疫学和细胞免疫学方面的培训。 与转录组学分析、成像和小鼠遗传学的相互作用。我将利用广泛的 洛克菲勒大学的资源，丹尼尔穆西达博士的指导，加布里埃尔维克多拉博士和咨询 委员会团队将在项目和职业发展的关键方面提供专业知识。结束时 在辅导阶段，我将配备必要的工具，在交叉点进行全面的研究 细胞通讯和免疫反应的独立研究员。

Project summary The highly specialized intestinal immune system is charged with maintaining tolerance to harmless stimuli from food and commensal bacteria, while providing protective immunity against pathogens. Dysregulation of this critical balance can lead to food allergy, inflammatory bowel disease (IBD), or increased susceptibility to enteric pathogens. Dendritic cells (DCs) are key players in intestinal homeostasis, finely orchestrating immune responses by presenting luminal antigens and inducing functional differentiation of CD4+ T cells into regulatory or pro-inflammatory subsets. The cellular mechanisms underlying the decision between tolerance or immunity to intestinal antigens remain unknown, largely because the identification and characterization of the exact DCs involved in these processes has been a decades-long technical challenge. To overcome this problem, I established the use of the LIPSTIC (Labeling Immune Partnerships by SorTagging Intercellular Contacts) technology in the gut, which enables proximity-dependent labeling of specific intestinal cell-cell interactions. The main goal of this research proposal is to elucidate the mechanisms that determine whether DCs will induce regulatory or pro-inflammatory T cells in vivo, by combining the LIPSTIC system with gene targeting, functional imaging, intersectional genetics, and interaction-based transcriptomics approaches. This proposal tests the hypothesis that DCs specialized for different functions exist in the intestine and that continuous dynamic reprogramming of DCs subsets by luminal content dictate tolerance or immunity to food and microbes. Specifically, this concept will be tested in mice under physiological (tolerance to food and microbes) and pathological (allergic sensitization, enteric infection, and colitis) scenarios. To this end, the K99 mentored phase (Aim 1) will reveal the exact nature of the DCs that induce tolerance or inflammation to food and the cellular and molecular mechanisms by which food-specific pTregs maintain oral tolerance. Next, the R00 independent phase (Aim 2) will focus on studying immune responses to commensals, with relevance to disorders such as colitis. Elucidating the DC populations, location and molecular mechanisms involved in tolerance to distinct microbes will reveal the nature of host-microbe interactions in health and disease. Together, this research program will lend fundamental insight into the etiology of certain intestinal disorders and will provide foundation for the development of new therapies for food allergy, colitis, and enteric infections, with profound implications for public health. The proposed development plan complements my training in mucosal immunology and cellular interactions with transcriptomics analysis, imaging, and mouse genetics. I will take advantage of the extensive resources of the Rockefeller University, the mentorship of Dr. Daniel Mucida, Dr. Gabriel Victora and the Advisory Committee team that will lend expertise in key aspects of the project and career development. At the end of the mentored phase, I will be equipped with the necessary tools to conduct comprehensive studies at the intersection of cellular communication and immune response to commensals as an independent investigator.