Innate and adaptive Treg in Immune tolerization of RA

RA 免疫耐受中的先天性和适应性 Treg

• 批准号: 6781373
• 负责人: Salvatore Albani
• 金额: $ 23.04万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-15 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6781373
• 关键词: CD antigens; CD28 molecule; CD4 molecule; T lymphocyte; biomarker; cell population study; cellular immunity; chemokine receptor; clinical research; flow cytometry; gene expression; human tissue; immune tolerance /unresponsiveness; immunogenetics; interleukin 10; interleukin 4; rheumatoid arthritis; synovial fluid; transforming growth factors

DESCRIPTION (provided by applicant): It has been suggested that two categories of Treg can be identified by phenotypical and functional characteristics. "Innate" and "adaptive" Treg would cooperate in limiting potentially noxious inflammatory processes. This regulatory function may be impaired in autoimmunity. Its restoration could provide novel therapeutic approaches. This project aims to unravel the role, which may pertain to Treg function in induction of tolerance to an antigenic peptide (dnaJP1) in rheumatoid arthritis (RA). 105 samples from RA patients treated in the context of a Phase 1 (completed) and a Phase II (ongoing) clinical trial with dnaJP1 or placebo have already been collected at the beginning of treatment and at monthly intervals. We will test whether mucosal tolerization to a peptide is associated with emergence of cells with a regulatory phenotype. We will also determine if a proportion of Treg are dnaJP1-specific, and if numbers and functional characteristics of these cells change as a consequence of immunotherapy. The specific aims are: Specific Aim 1: To characterize "innate" and "adaptive" Treg in serial samples obtained from a tolerization trial in rheumatoid arthritis and to explore their functional role in the tolerization process. SA1a: Peripheral blood mononuclear cells (PBMC), synovial fluid mononuclear cells (SFMC) and T cells obtained from synovial membranes of RA patients will be evaluated by FACS analysis for phenotypical markers characteristic of T regulatory T cells. In particular, levels of CD25, CD4, CTLA4 and CCR4 will be studied. "Innate" and "adaptive" T cells will be differentiated based on a set of phenotypical and functional variables, including levels of CD25 expression and quantification on sorted cells by real time PCR (TaqMan) gene expression of several molecules putatively involved in Treg function. These genes will include IL-10, TGF beta, IL-4, FOXP3, CTLA. Samples obtained will be also tested in in vitro studies to explore regulatory properties of "innate" and "adaptive" Treg on T cell responses to recall antigens as well as to antigens (gp39, dnaJP1 and p205) putatively involved in the pathogenic process. SA1b: Clinical information will be compared with immunological data Specific Aim 2: To investigate whether some Treg have specificity for dnaJpl and whether functional and phenotypical characteristics of dnaJP1-specific Treg are associated with the course of the tolerization process. Functional characteristics of sorted cells will be evaluated by TaqMan.

描述（由申请人提供）：有人建议可以通过表型和功能特征来鉴定两类 Treg。 “先天”和“适应性”Treg 将合作限制潜在有害的炎症过程。这种调节功能可能会因自身免疫而受损。它的修复可以提供新的治疗方法。该项目旨在揭示 Treg 在诱导类风湿性关节炎 (RA) 抗原肽 (dnaJP1) 耐受中的作用，这可能与 Treg 功能有关。在 dnaJP1 或安慰剂的 1 期（已完成）和 2 期（正在进行）临床试验中，已在治疗开始时和每月间隔收集了 105 份 RA 患者的样本。我们将测试粘膜对肽的耐受是否与具有调节表型的细胞的出现相关。我们还将确定一部分 Treg 是否具有 dnaJP1 特异性，以及这些细胞的数量和功能特征是否因免疫治疗而发生变化。具体目标是： 具体目标 1：表征从类风湿性关节炎耐受试验中获得的系列样本中的“先天”和“适应性”Treg，并探索它们在耐受过程中的功能作用。 SA1a：将通过 FACS 分析评估外周血单核细胞 (PBMC)、滑液单核细胞 (SFMC) 和从 RA 患者滑膜获得的 T 细胞，以了解 T 调节性 T 细胞的表型标志物特征。特别是，将研究 CD25、CD4、CTLA4 和 CCR4 的水平。 “先天性”和“适应性”T 细胞将根据一组表型和功能变量进行区分，包括 CD25 表达水平以及通过实时 PCR (TaqMan) 推测参与 Treg 功能的几种分子基因表达对分选细胞进行的定量。这些基因包括 IL-10、TGF beta、IL-4、FOXP3、CTLA。获得的样本还将在体外研究中进行测试，以探索“先天”和“适应性”Treg 对召回抗原以及推测参与致病过程的抗原（gp39、dnaJP1 和 p205）的 T 细胞反应的调节特性。 SA1b：将临床信息与免疫学数据进行比较具体目标2：研究某些Treg是否对dnaJP1具有特异性以及dnaJP1特异性Treg的功能和表型特征是否与耐受过程的过程相关。 TaqMan 将评估分选细胞的功能特征。