Keratinocyte regulation of skin immunity

描述（由申请人提供）： 假设：本研究的目的是确定KC如何控制皮肤中T细胞介导的免疫。我们将利用人类和小鼠模型的常见皮肤病，过敏性接触性皮炎（ACD）。我们选择这种皮肤状况，因为它是退伍军人常见的皮肤病。我们假设角质形成细胞（KC）是自然杀伤（NK）T细胞的致耐受性抗原呈递细胞。KC-NKT-细胞通过CD 1d相互作用诱导NKT克隆无能，在维持外周耐受中起重要作用。我们将测试我们的中心假设KC上的CD 1d可能耐受亲表皮性NKT，可能抑制接触性皮炎。我们将研究KC-NKT-细胞的相互作用，以及它们如何影响皮肤中细胞介导的免疫力，使用小鼠模型以及人类KC和NKT细胞的直接研究。具体目标：有三个具体目标：1）研究体外KC-NKT-细胞相互作用的作用以及这是否耐受NKT-细胞。2)确定无反应性iNKT细胞在小鼠ACD的传入和传出阶段中的作用。3)确定表皮KC的CD 1d丢失对ACD（耐受丧失）的影响。与VA的相关性：接触性皮炎是工业化国家非常常见的炎症性皮肤病。这是患者的主要健康问题，对经济有重大影响。接触性皮炎也是退伍军人的一个重要问题，这是退伍军人访问皮肤科诊所的常见原因。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。将使用动物模型系统在移植有来自CD 1d基因敲除小鼠的皮肤的小鼠中进行ACD。这种体内模型将使我们能够确认KC衍生的CD 1d的作用，并将扩展我们在体内控制先天性免疫的CD 1d的作用的体外研究。操作步骤：KC、NKT细胞、单核细胞的组织培养、细胞增殖试验、流式细胞术、ELISA、qPCR、基因靶向小鼠的体内研究将用于解决本提案的中心焦点问题：KC CD 1d如何控制NKT细胞反应，从而控制ACD的过程。结果对退伍军人事务部的意义：皮炎是退伍军人中常见的皮肤病，就像在一般人群中一样。从中东冲突返回的退伍军人将经历健康问题，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质，将进一步了解先天免疫（NKT细胞）与与环境界面（在本提案中为皮肤中的上皮表面）之间的界面，并与其他器官中的过敏性疾病（如哮喘）相关。拟议的免疫学研究将利用ACD作为模型来剖析这种疾病的病理机制。这些研究将是非常有用的了解这种常见的皮肤病，并将适用于过敏性健康问题的退伍军人从中东冲突返回。 公共卫生相关性： 对退伍军人医疗保健的潜在影响：皮炎是退伍军人中常见的皮肤病，因为它是在一般人群中。从中东冲突返回的退伍军人将经历健康问题，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质，将进一步了解先天免疫（NKT细胞）与与环境界面（在本提案中为皮肤中的上皮表面）之间的界面，并与其他器官中的过敏性疾病（如哮喘）相关。拟议的免疫学研究将利用ACD作为模型来剖析这种疾病的病理机制。这些研究将是非常有用的了解这种常见的皮肤病，并将适用于过敏性健康问题的退伍军人从中东冲突返回。

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. PUBLIC HEALTH RELEVANCE: Potential Impact on Veteran's Healthcare: 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.