Role of JNK2 and TLR6 during Y. enterocolitica induced mucosal immune responses

JNK2 和 TLR6 在小肠结肠炎耶尔森氏菌诱导的粘膜免疫反应中的作用

• 批准号: 7755830
• 负责人: R William DePaolo
• 金额: $ 12.71万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7755830
• 关键词: Antigen-Presenting Cells; Antigens; Biological Models; Breeding; CD4 Positive T Lymphocytes; Calcium; Cells; Gastroenteritis; Generations; Goals; Immune; In Vitro; Infection; Inflammation; Ingestion; Interleukin-10; Intestinal Mucosa; Intestines; Knockout Mice; MAP Kinase Gene; MAPK8 gene; MAPK9 gene; Mediating; Mesenteric Lymphadenitis; Molecular; Monoclonal Antibodies; Mucosal Immune Responses; Mus; Oral; Pathogenesis; Peripheral; Play; Principal Investigator; Reporter; Research; Role; Site; Small Intestines; Structure of aggregated lymphoid follicle of small intestine; T-Lymphocyte; T-Lymphocyte Subsets; TLR6 gene; Tissues; Transgenic Organisms; Tropism; Virulence Factors; Yersinia; Yersinia enterocolitica; Yersinia pestis; bacterial lysate; cytokine; insight; mucosal site; mutant; programs; public health relevance; response

DESCRIPTION (provided by applicant): IL-10 is a critical cytokine for homeostatic responses in the intestinal mucosa, both against orally ingested antigens and against commensal microflora. IL-10 can exert its effects through the induction of DC that prime tolerogenic responses, referred to as tolerogenic DC. It is proposed that tolerogenic DC prime type-1 regulatory (Tr1) cells, a subset of T cell that produce high amounts of IL-10 and suppress inflammation. However, the molecular mechanism of tolerogenic DC and (Tr1) cell generation in the intestine remain poorly understood. The goal of this proposal is to determine how IL-10-producing cells are generated in the intestinal immune tissue using Y. enterocolitica infection as a model system. What makes Y. enterocolitica an ideal choice for examining induction of IL-10-mediated mucosal responses is because of its tropism for immune sites of the small intestine, specifically the Peyer's patches and the MLN, both sites of immune cell induction and T cell priming. More importantly, Y. enterocolitica harbors the Low Calcium Response V Antigen (LcrV) an essential virulence factor found in all three pathogenic Yersinia species and which has been shown to induce IL-10 from antigen presenting cells (APC). Using systemic Y. pestis infection we found that LcrV induced tolerogenic DC and primed a Tr1 cell response in a TLR6 dependent manner. Furthermore, our initial studies suggest that TLR6 is uniquely involved in the generation of tolerogenic DC and Tr1 through activation of the MAPK JNK. We hypothesize that TLR6 and JNK will play a key role in the generation of mucosal intestinal tolerogenic DC and Tr1 cells. Furthermore, we anticipate that TLR6 and JNK play an important role in Y. enterocolitica pathogenesis. Our specific aims will investigate [sic], 1) Evaluate the induction of tolerogenic DC and Tr1 cells using mucosal DC from TLR-deficient and JNK-deficient mice stimulated in vitro by LcrV or infected with Y. enterocolitica. 2) Evaluate the role TLR6 and JNK play during pathogenesis of oral Y. enterocolitica infection. In this aim we will utilize knockout mice as well as by using IL-10-T cell reporter mice bred to TLR-deficient and JNK-deficient mice in hopes to identify the mucosal sites of IL-10 induction and evaluate whether mucosal CD4 T cells induced during infection can transfer tolerance and spread tolerance to a second antigen. 3) Use monoclonal antibodies against LcrV and LcrV deletion mutants to identify specific residues within LcrV responsible for inducing IL-10 and activating JNK. PUBLIC HEALTH RELEVANCE: Understanding the molecular mechanism that Y. enterocolitica induces intestinal Tr1 and tolerogenic DC, may give us critical insight into understanding how intestinal tissue generates regulatory responses homeostatically.

描述（由申请人提供）： IL-10 是肠粘膜稳态反应的关键细胞因子，无论是针对口服抗原还是针对共生微生物群。 IL-10 可以通过诱导引发耐受性反应的 DC（称为耐受性 DC）来发挥其作用。据推测，耐受性 DC Prime 1 型调节 (Tr1) 细胞是 T 细胞的一个子集，可产生大量 IL-10 并抑制炎症。然而，肠道中耐受性 DC 和 (Tr1) 细胞生成的分子机制仍知之甚少。该提案的目标是使用小肠结肠炎耶尔森氏菌感染作为模型系统，确定肠道免疫组织中如何产生 IL-10 细胞。小肠结肠炎耶尔森氏菌之所以成为检查 IL-10 介导的粘膜反应诱导的理想选择，是因为它对小肠免疫位点的趋向性，特别是派尔氏集结和 MLN，这两个位点都是免疫细胞诱导和 T 细胞启动的位点。更重要的是，小肠结肠炎耶尔森氏菌含有低钙反应 V 抗原 (LcrV)，这是一种在所有三种致病性耶尔森氏菌中发现的重要毒力因子，并且已被证明可以诱导抗原呈递细胞 (APC) 产生 IL-10。使用全身性鼠疫耶尔森氏菌感染，我们发现 LcrV 诱导耐受性 DC 并以 TLR6 依赖性方式引发 Tr1 细胞反应。此外，我们的初步研究表明，TLR6 通过激活 MAPK JNK 独特地参与耐受性 DC 和 Tr1 的生成。我们假设 TLR6 和 JNK 将在粘膜肠道耐受性 DC 和 Tr1 细胞的生成中发挥关键作用。此外，我们预计 TLR6 和 JNK 在小肠结肠炎耶尔森氏菌发病机制中发挥重要作用。我们的具体目标将研究[原文如此]，1) 使用来自 LcrV 体外刺激或感染小肠结肠炎耶尔森氏菌的 TLR 缺陷和 JNK 缺陷小鼠的粘膜 DC 来评估耐受性 DC 和 Tr1 细胞的诱导。 2)评估TLR6和JNK在口腔小肠结肠炎耶尔森氏菌感染发病机制中的作用。为此，我们将利用基因敲除小鼠以及与 TLR 缺陷和 JNK 缺陷小鼠交配的 IL-10-T 细胞报告小鼠，希望鉴定 IL-10 诱导的粘膜位点，并评估感染过程中诱导的粘膜 CD4 T 细胞是否可以转移耐受性并将耐受性传播到第二种抗原。 3) 使用针对 LcrV 和 LcrV 缺失突变体的单克隆抗体来鉴定 LcrV 内负责诱导 IL-10 和激活 JNK 的特定残基。 公共卫生相关性：了解小肠结肠炎耶尔森氏菌诱导肠道 Tr1 和耐受性 DC 的分子机制，可能会让我们深入了解肠道组织如何产生稳态调节反应。