Characterization of Innate immune receptors

先天免疫受体的表征

• 批准号: 7451469
• 负责人: DETLEF SCHUPPAN
• 金额: $ 25.5万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7451469
• 关键词: Affect; Affinity Chromatography; African; Amino Acids; Antigens; Autoantigens; Autoimmune Responses; Autoimmune thyroiditis; Autoimmunity; Bacterial Polysaccharides; Binding; Blocking Antibodies; CCL2 gene; Celiac Disease; Cell membrane; Cells; Cereals; Complication; Data; Dendritic Cells; Diet; Disease; Enzymes; Epithelial; Epithelial Cells; European; Exclusion; Genetic Predisposition to Disease; Gliadin; Glutamates; Glutamine; Gluten; Goals; Human; IL8 gene; Immune System Diseases; Immune response; Immunity; Immunoglobulin A; Immunologic Receptors; Individual; Individual Differences; Inflammatory; Ingestion; Insulin-Dependent Diabetes Mellitus; Interferons; Interleukin-8; Intestinal Diseases; Intestinal Mucosa; Intestines; Label; Lead; Link; Lipopolysaccharides; MALDI-TOF Mass Spectrometry; Malabsorption Syndromes; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Minor; Natural Immunity; Pathogenesis; Pathway interactions; Patients; Pattern recognition receptor; Peptide Receptor; Peptides; Personal Satisfaction; Production; Proteins; Proteomics; Public Health; Reaction; Receptor Cell; Regulation; Signal Pathway; Small Interfering RNA; Small Intestines; Symptoms; System; T-Cell Activation; T-Lymphocyte; Techniques; Therapeutic; Tissues; Transglutaminases; Ursidae Family; Wheat; base; cell mediated immune response; gastrointestinal; glutenin; inhibitor/antagonist; novel; receptor; receptor function; response; viral RNA

DESCRIPTION (provided by applicant): Celiac disease (Cd) is a T cell mediated disease of the small intestine. It is triggered by dietary gluten proteins from cereals and affects 1 in 130 US citizens. While Cd can lead to severe malabsorption, most patients present with minor or atypical (nondiarrheal) symptoms. Cd is associated with secondary autoimmunity, such as type 1 diabetes or autoimmune thyroiditis, and longterm Cd that has not been treated with a strict gluten free diet can lead to (gastrointestinal) malignancy. Gluten peptides that have been deamidated by the Cd autoantigen tissue transglutaminase bind strongly to HLA-DQ2 or -DQ8, the essential genetic predisposition for Cd. This results in intestinal Th1 T cell activation and mucosal destruction (adaptive immunity). Apart from triggering adaptive immunity, recent data suggest that gluten (gliadin) can also stimulate innate immunity, i.e., the immediate and relatively nonspecific immune response to common foreign antigens, such as bacterial polysaccharide or viral RNA. The variable contribution of innate immunity to gliadin could explain why only 2- 5% of those individuals that carry HLA-DQ2 or -DQ8 finally develop Cd. The responsible gliadin peptide(s) and the receptors mediating innate immunity to gluten remain to be identified. Our preliminary results show that a peptic-tryptic digest of gliadin which contains roughly 1000 different gliadin peptides triggers a marked innate immune response in human dendritic but also intestinal epithelial cells, as assessed by release of the inflammatory mediators interleukin 8 and MCP-1. Based on these data we plan to: 1. Identify the gliadin peptide(s) that drive innate immunity after their chromatographic separation by using dendritic (intestinal epithelial) as indicators and MALDI-TOF mass spectrometry. 2. Isolate and characterize the innate immune receptor(s) on these cells that trigger(s) the innate immune response to gliadin by use of affinity chromatography of labeled cell membranes on the identified immobilized stimulatory gliadin peptide(s) and MALDI-TOF mass spectrometry. Identity and function of the receptor(s) will further be confirmed by use of function blocking antibodies, siRNA and signaling pathway inhibitors. We anticipate that identification of the receptor(s) responsible for the innate immune response to gliadin will lead to a better understanding of the pathogenesis of Cd, and possibly to novel nondietary therapies to treat this common intestinal disorder. PUBLIC HEALTH RELEVANCE Celiac disease (Cd) is a common immune disease of the small intestine that affects 1 in 130 US citizens. It is triggered by ingestion of the storage proteins of wheat (glutenins and especially gliadins) and related cereals. Cd that is not treated by strict dietary gluten exclusion can lead to severe malabsorption and malignancy, and is associated with secondary autoimmunity, such as type 1 diabetes. While the adaptive (T cell mediated) immune response to gluten peptides that leads to destruction of the resorptive intestinal mucosa is well understood, there is a yet ill defined innate immune response (the immediate and relatively nonspecific reaction to foreign antigens, such as bacterial polysaccharide or viral RNA) to gliadins. The responsible gliadin peptide(s) and the receptors mediating innate immunity to gluten remain to be identified. We plan to identify the gliadin peptide(s) that drive innate immunity in human dendritic and intestinal epithelial cells and characterize their responsive innate immune receptor(s) by use of proteomic techniques, affinity chromatography and functional studies. We anticipate that identification of the receptor(s) responsible for the innate immune response to gliadin will lead to a better understanding of the pathogenesis of Cd, and possibly to novel nondietary therapies to treat this common intestinal disorder.

描述（由申请人提供）：乳糜泻（Cd）是一种 T 细胞介导的小肠疾病。它是由谷物中的膳食麸质蛋白引发的，每 130 名美国公民中就有 1 人受到影响。虽然镉可导致严重吸收不良，但大多数患者会出现轻微或非典型（非腹泻）症状。镉与继发性自身免疫有关，例如 1 型糖尿病或自身免疫性甲状腺炎，未经严格无麸质饮食治疗的长期镉可导致（胃肠道）恶性肿瘤。已被 Cd 自身抗原组织转谷氨酰胺酶脱酰胺的麸质肽与 HLA-DQ2 或 -DQ8（Cd 的基本遗传倾向）强烈结合。这会导致肠道 Th1 T 细胞激活和粘膜破坏（适应性免疫）。除了触发适应性免疫外，最近的数据表明，麸质（麦醇溶蛋白）还可以刺激先天免疫，即对常见外来抗原（例如细菌多糖或病毒 RNA）的直接且相对非特异性的免疫反应。先天免疫对麦醇溶蛋白的不同贡献可以解释为什么携带 HLA-DQ2 或 -DQ8 的个体中只有 2-5% 最终出现 Cd。负责的麦醇溶蛋白肽和介导对麸质先天免疫的受体仍有待鉴定。 我们的初步结果表明，通过炎症介质白细胞介素 8 和 MCP-1 的释放来评估，含有大约 1000 种不同麦醇溶蛋白肽的麦醇溶蛋白的消化胰蛋白酶消化物会在人类树突细胞和肠上皮细胞中引发显着的先天免疫反应。基于这些数据，我们计划： 1. 通过使用树突（肠上皮）作为指示剂和 MALDI-TOF 质谱法，在色谱分离后鉴定驱动先天免疫的麦醇溶蛋白肽。 2. 通过在已识别的固定刺激性麦醇溶蛋白肽和 MALDI-TOF 质量上使用标记细胞膜的亲和层析，分离并表征这些细胞上触发对麦醇溶蛋白的先天免疫反应的先天免疫受体光谱测定法。受体的身份和功能将通过使用功能阻断抗体、siRNA 和信号通路抑制剂进一步确认。 我们预计，对麦醇溶蛋白的先天免疫反应负责的受体的鉴定将有助于更好地了解镉的发病机制，并可能开发出新的非饮食疗法来治疗这种常见的肠道疾病。 公共卫生相关性 乳糜泻 (Cd) 是一种常见的小肠免疫疾病，每 130 名美国公民中就有 1 人受到影响。它是由摄入小麦（麦谷蛋白，尤其是麦醇溶蛋白）和相关谷物的储存蛋白引发的。未经严格饮食麸质排除治疗的镉可导致严重吸收不良和恶性肿瘤，并与继发性自身免疫相关，例如 1 型糖尿病。虽然对谷蛋白肽的适应性（T 细胞介导的）免疫反应导致肠粘膜再吸收的破坏已被充分了解，但尚不清楚的先天免疫反应（对外来抗原（例如细菌多糖）的直接且相对非特异性的反应）或病毒RNA）至麦醇溶蛋白。负责的麦醇溶蛋白肽和介导对麸质先天免疫的受体仍有待鉴定。我们计划鉴定在人类树突状细胞和肠上皮细胞中驱动先天免疫的麦醇溶蛋白肽，并通过使用蛋白质组学技术、亲和层析和功能研究来表征其反应性先天免疫受体。我们预计，对麦醇溶蛋白的先天免疫反应负责的受体的鉴定将有助于更好地了解镉的发病机制，并可能开发出新的非饮食疗法来治疗这种常见的肠道疾病。