Keratinocyte regulation of skin immunity

描述(由申请人提供)： 假设：这项研究的目的是确定KC如何控制皮肤中T细胞介导的免疫。我们将利用常见皮肤病--过敏性接触性皮炎(ACD)的人类和小鼠模型。我们选择这种皮肤病是因为它是退伍军人的一种常见皮肤病。我们假设角质形成细胞(KC)是自然杀伤(NK)T细胞的耐受抗原提呈细胞。KC-NKT-细胞通过CD1d相互作用，通过诱导NKT克隆性无能，在维持外周免疫耐受中发挥重要作用。我们将验证我们的中心假说CD1d对KC可能耐受嗜表性NKT，潜在地抑制接触性皮炎。我们将使用小鼠模型以及对人类KC和NKT-细胞的直接研究，研究KC-NKT-细胞之间的相互作用，以及它们如何影响皮肤中的细胞免疫。具体目的：有三个具体目标：1)研究KC-NKT-细胞相互作用的体外效应及其对NKT-细胞的耐受性。2)明确无能iNKT细胞在ACD传入和传出过程中的作用。3)探讨表皮KC缺失CD1d对耐受性丧失的影响。与退伍军人事务部的相关性：接触性皮炎是工业化世界中非常常见的炎症性皮肤病。它是患者的一个主要健康问题，对经济有重大影响。接触性皮炎对退伍军人来说也是一个重大问题，是退伍军人去皮肤科诊所就诊的常见原因。受试者群体：KC和NKT细胞将从健康对照中获得，用于体外研究。一种动物模型系统将用于移植CD1d基因敲除小鼠皮肤的小鼠的ACD。这一体内模型将使我们能够确认KC来源的CD1d的作用，并将扩大我们对CD1d在体内控制天然免疫的作用的体外研究。程序：KC的组织培养，NKT细胞，单核细胞，细胞增殖分析，流式细胞仪，ELISA，qPCR，基因靶向小鼠的体内研究将被用来解决这项提议的中心焦点问题：KC CD1d如何控制NKT细胞的反应，这控制了ACD的进程。研究结果对退伍军人的意义：皮炎是退伍军人中常见的皮肤病，在普通人群中也是如此。从中东冲突归来的退伍军人将遇到健康问题，包括影响皮肤的免疫系统疾病，如接触性皮炎。拟议的研究具有基础性，将进一步了解先天免疫(NKT细胞)和与环境(在本建议中为皮肤上皮表面)相互作用的组织之间的界面，并与其他器官的过敏性疾病(如哮喘)有关。拟议的免疫学研究将以ACD为模型来剖析该病的发病机制。这些研究将对了解这种常见的皮肤病非常有用，并将应用于从中东冲突归来的退伍军人的过敏性健康问题。

DESCRIPTION (provided by applicant): Hypothesis: The purpose of this study is to define how KC control T-cell mediated immunity in the skin. We will utilize both human and mouse models of the common skin condition, allergic contact dermatitis (ACD). We selected this skin condition because it is a common skin disease in Veterans. We hypothesize that Keratinocytes (KC) are tolerigenic Antigen presenting cells for Natural killer (NK)T-cells. KC-NKT-cell interactions via CD1d plays an important role in maintaining peripheral tolerance by inducing NKT clonal anergy. We will test our central hypothesis CD1d on KC may tolerize epidermotropic NKT, potentially dampening contact dermatitis. We will study KC-NKT-cell interactions, and how they affect cell mediated immunity in the skin, using mouse models as well as direct studies of human KC and NKT-cells. Specific Objectives: There are three specific objectives: 1) To study the effects of KC- NKT-cell interactions in vitro and whether this tolerizes NKT-cells. 2) To define the role of anergic iNKT-cells in the afferent and efferent phases of murine ACD. 3) To determine the effects of the loss of CD1d by epidermal KC on ACD (loss of tolerance). Relevance to the VA: Contact dermatitis a very common inflammatory skin diseases in the industrialized world. It is a major health concern for patients and has a major impact on the economy. Contact dermatitis is a significant problem for veterans as well, representing a common cause for visits to Dermatology clinics in veterans. Subject Populations: KC and NKT-cells will be obtained from healthy controls for in vitro studies. An animal model system will be used to ACD in mice engrafted with skin from CD1d gene knockout mice. This in vivo model will allow us to confirm the role of KC derived CD1d, and will extend our in vitro studies on the role of CD1d in controlling innate immunity in vivo. Procedures: Tissue culture of KC, NKT-cells, monocytes, cell proliferation assays, flow cytometry, ELISA, qPCR, in vivo studies with gene targeted mice will be utilized to address the questions that are the central focus of this proposal: How KC CD1d controls NKT-cell responses, and this controls the course of ACD. Significance of findings to the VA: Dermatitis a common skin disease in Veterans as it is in the general population. Veterans returning from the Middle East Conflicts will experience health problems, including diseases of the immune system that will affect the skin, such as contact dermatitis. The proposed studies are of a fundamental nature that will further the understanding of the interface between innate immunity (NKT-cells), and tissues that interface with the environment (in this proposal epithelial surfaces in the skin), and are relevant to allergic diseases in other organs such as asthma. The proposed immunological study will utilize ACD as a model to dissect out patho-mechanisms of this disease. These studies will be of great utility in understanding this common skin disease, and will be applied to allergic health problems in Veterans returning from the Middle East Conflicts.