Generation and characterization of intestinal CD8+ regulatory T cell lines

肠道 CD8 调节性 T 细胞系的产生和表征

• 批准号: 7821735
• 负责人: Lloyd F Mayer
• 金额: $ 49.7万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7821735
• 关键词: Address; Adult; Adverse effects; Applications Grants; Area; Autoantibodies; Autoimmune Diseases; Autoimmunity; Automobile Driving; Biochemical; CD4 Positive T Lymphocytes; CD8B1 gene; Cell Line; Cell Separation; Cell surface; Cells; Characteristics; Child; Chronic; Clinical Research; Crohn' s disease; Data; Defect; Development; Disease; Epithelial Cells; Equilibrium; Experimental Autoimmune Encephalomyelitis; Functional disorder; Funding; Gastrointestinal tract structure; Generations; Goals; Health; Homeostasis; Human; Immune response; Immune system; Immunohistochemistry; Immunology; Immunosuppressive Agents; In Vitro; Incidence; Individual; Inflammation; Inflammatory Bowel Diseases; Institutes; Insulin-Dependent Diabetes Mellitus; Intestines; Laboratories; Lamina Propria; Lead; Maintenance; Mediating; Methods; Microarray Analysis; Monoclonal Antibodies; Mucous Membrane; Multiple Sclerosis; Normal Cell; Patients; Peripheral; Pharmaceutical Preparations; Play; Population; Postdoctoral Fellow; Prevalence; Recovery; Relapse; Reporting; Research; Rodent; Role; Suppressor-Effector T-Lymphocytes; Surface; T-Lymphocyte; Techniques; Technology; Therapeutic; Therapeutic Clinical Trial; Tissues; Translating; Ulcerative Colitis; United States; Work; base; commensal microbes; disorder control; inhibitor/antagonist; novel; novel therapeutics; peripheral blood; precursor cell; prevent; public health relevance; tool

DESCRIPTION (provided by applicant): This grant application addresses broad Challenge Area (04): Clinical Research and specific challenge topic 04-AI-101: Develop novel methods and address key questions in mucosal immunology. The proposal draws from strong preliminary data from our laboratory documenting defects in the activation of CD8+ Tregs in vitro in patients with IBD (Crohn's disease and ulcerative colitis) and exciting new data detailing defective CD8+ Tregs ex vivo from CD mucosa (intestine). In this proposal we will characterize normal human CD8+ regulatory T cell lines (derived from the lamina propria), define their mechanism of suppression and develop approaches to expand precursors of these cells from the peripheral blood. Our preliminary data suggest that normal LP derived CD8+ Tregs are potent inhibitors of immune responses while similar lines derived from patients with Crohn's disease fail to suppress. If we can generate Treg lines from peripheral blood precursors from CD patients, we will have developed a novel cell based therapeutic for the treatment of CD. The first aim will focus on defining phenotypic and functional characteristics of normal lamina propria regulatory CD8+ T cell lines. This will include the development of monoclonal antibodies against surface molecules involved in mediating the suppressor effector function. This mAb will also allow for the identification of precursor cells in the PB. The second aim will compare and contrast LP CD8+ T cell lines from Crohn's and ulcerative colitis patients vs. normal controls. An important feature of this aim will be a microarray analysis that will not only identify specific differences in CD vs normal cell lines but also add a different perspective on the mechanism of suppression by normal lines. Finally, the third aim will focus on efforts to identify precursors of CD8+ Tregs in the PB and develop strategies to expand these cells ex vivo. The ultimate goal will be to utilize the peripheral blood derived cell lines as novel therapeutics in the treatment of IBD. Funding for these studies will allow us to hire two additional individuals (one post-doctoral fellow to perform the biochemical analysis of the cell surface molecule involved in suppressor effector function and one technician involved in the immunohistochemistry, cell sorting and analysis. The microarray studies in aim 2 will be performed at the Baylor Institute for Immunology Research where additional technicians will be required. These latter studies highlight a new alliance between Mount Sinai and the BIIR that provides enabling technologies to translate the results from these studies into novel cell based therapeutics for clinical trials. PUBLIC HEALTH RELEVANCE: Crohn's disease (CD) and ulcerative colitis (UC), the major forms of Inflammatory bowel disease (IBD), are a growing and serious health concern in the United States with reported prevalence rate as high as 43.6 per 100,000. IBD is characterized by chronic and relapsing inflammation of the gastrointestinal tract representing defects in the control of immune responses in the intestine. We believe that CD8+ regulatory cells are an important cell population involved in controlling local immune responses in the gut and propose to study this cell subset and develop tools that will allow for the use of this cell population as a potential therapy for CD.

描述（由申请人提供）：本拨款申请涉及广泛的挑战领域 (04)：临床研究和具体挑战主题 04-AI-101：开发新方法并解决粘膜免疫学中的关键问题。该提案借鉴了我们实验室的强有力的初步数据，记录了 IBD（克罗恩病和溃疡性结肠炎）患者体外 CD8+ Tregs 激活的缺陷，以及详细描述 CD 粘膜（肠）离体 CD8+ Tregs 缺陷的令人兴奋的新数据。在本提案中，我们将描述正常人 CD8+ 调节性 T 细胞系（源自固有层），定义其抑制机制，并开发从外周血中扩增这些细胞前体的方法。我们的初步数据表明，正常 LP 衍生的 CD8+ Tregs 是免疫反应的有效抑制剂，而来自克罗恩病患者的类似细胞系则无法抑制。如果我们能够从 CD 患者的外周血前体中产生 Treg 系，我们将开发出一种新的基于细胞的治疗 CD 的疗法。第一个目标将集中于定义正常固有层调节性 CD8+ T 细胞系的表型和功能特征。这将包括开发针对参与介导抑制效应功能的表面分子的单克隆抗体。该 mAb 还可用于鉴定 PB 中的前体细胞。第二个目标是比较和对比来自克罗恩病和溃疡性结肠炎患者与正常对照的 LP CD8+ T 细胞系。这一目标的一个重要特征是微阵列分析，它不仅可以识别 CD 与正常细胞系的具体差异，而且还可以从不同的角度来了解正常细胞系的抑制机制。最后，第三个目标将集中于鉴定 PB 中 CD8+ Tregs 的前体，并制定离体扩增这些细胞的策略。最终目标是利用外周血来源的细胞系作为治疗 IBD 的新疗法。这些研究的资金将使我们能够雇用另外两名人员（一名博士后研究员对参与抑制效应功能的细胞表面分子进行生化分析，一名技术人员参与免疫组织化学、细胞分选和分析。目标 2 中的微阵列研究将在贝勒免疫学研究所进行，那里需要更多的技术人员。这些后面的研究强调了西奈山和 BIIR 提供使能技术，将这些研究的结果转化为用于临床试验的新型细胞疗法。 公共卫生相关性：克罗恩病 (CD) 和溃疡性结肠炎 (UC) 是炎症性肠病 (IBD) 的主要形式，在美国是一个日益严重的健康问题，据报告患病率高达每 100,000 人 43.6 人。 IBD 的特征是胃肠道慢性和复发性炎症，代表肠道免疫反应控制缺陷。我们认为 CD8+ 调节细胞是参与控制肠道局部免疫反应的重要细胞群，并建议研究该细胞亚群并开发工具，以允许使用该细胞群作为 CD 的潜在疗法。