REGULATION OF AUTOIMMUNITY WITH T-CELL RECEPTOR PEPTIDES

T 细胞受体肽调节自身免疫

• 批准号: 2264749
• 负责人: ARTHUR A. VANDENBARK
• 金额: $ 24.82万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-08-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2264749
• 关键词: SCID mouse; T cell receptor; T lymphocyte; antireceptor antibody; autoimmunity; clone cells; epitope mapping; experimental allergic encephalomyelitis; human subject; immunotherapy; monoclonal antibody; multiple sclerosis; myelin basic proteins; passive immunization; polymerase chain reaction; synthetic peptide; tissue /cell culture

The overall goal of this proposal is to test the hypothesis that induction of immunity against synthetic peptides that resemble natural human TCR V-beta-5.2 idiotopes can regulate potentially pathogenic T cells that preferentially express V-beta-5.2. Our prior studies in rats and mice demonstrated that TCR V-beta-8.2 CDR2 peptides can prevent and treat clinical signs of experimental encephalomyelitis by inducing T cells and antibodies that can inhibit the mono-V-beta-8.2 T cell response to myelin basic protein (BP). Recently, we found biased use of V-beta-5.2 and V-beta-6.1 by BP-specific T cells from multiple sclerosis (MS) patients, a result corroborated by others who found V-beta-5.2+ T cells with a BP-specific CDR3 motif in MS plaques. We have now shown that anti-TCR specific T cells and antibodies can be induced by injecting TCR V-beta-5.2 or V-beta-6.1 peptides into MS patients. Thus the stage is set to test whether these T cells and antibodies can regulate V-beta-5.2 + T cells, including those specific for BP. Successful regulation of BP reactivity would allow a critical evaluation of the role of BP in the MS disease process, and would establish a prototypic approach for the treatment of other autoimmune diseases characterized by limited V gene expression. AIM 1. To assess the frequency, specificity, TCR repertoire, and encephalitogenicity of human BP-specific T cells. In this aim, we will develop the ability to rapidly and repeatedly evaluate MS disease- associated changes in BP-specific T cell responses in blood and CSF. Moreover, we will assess the encephalitogenic activity of these T cells by passive transfer into MHC compatible, bone marrow reconstituted SCID-Hu mice. AIM 2. To establish the effects of TCR peptide injection on TCR and BP responses. With a focus on V-beta-5.2, we will inject overlapping peptides into MS patients with V-beta-5.2-biased responses to BP to determine which regions are immunodominant T and B cell idiotopes. In responders, we will monitor functional changes in V-beta-5.2+ T cells and in responses to BP. AIM 3. To evaluate potential regulatory mechanisms induced by TCR peptides. In this aim, we will isolate and characterize distinct anti-TCR peptide-specific T cell clonotypes and antibodies induced after TCR peptide boosting. We will evaluate each T cell clonotype and affinity purified antibodies for their ability to regulate autologous V-beta-5.2+ BP-reactive T cells in vitro and in the SCID-Hu mouse.

本提案的总体目标是检验以下假设： 诱导针对类似于天然合成肽的免疫 人TCRV-β-5.2独特表位可调节潜在致病性T细胞 优先表达V-β-5.2的细胞。我们之前在老鼠身上的研究 和小鼠证明TCR V-β-8.2 CDR 2肽可以预防和 通过诱导T治疗实验性脑脊髓炎的临床体征 可以抑制单V-β-8.2 T细胞应答的细胞和抗体 髓鞘碱性蛋白（BP）。 最近，我们发现， 来自多发性硬化的BP特异性T细胞的V-β-5.2和V-β-6.1 (MS)患者，这一结果得到了其他发现V-β-5.2 + T 在MS斑块中具有BP特异性CDR 3基序的细胞。 我们现在已经展示了 抗TCR特异性T细胞和抗体可以通过注射 TCR V-β-5.2或V-β-6.1肽的免疫调节。 因此， 将测试这些T细胞和抗体是否可以调节 V-β-5.2 + T细胞，包括BP特异性T细胞。 成功 BP反应性的调节将允许对 BP在MS疾病过程中的作用，并将建立一个原型 治疗其他自身免疫性疾病的方法， 限制V基因表达。 AIM 1.为了评估频率，特异性，TCR库，和 人BP特异性T细胞的致脑炎性。 为此，我们会 开发快速和重复评估MS疾病的能力- 血液和CSF中BP特异性T细胞应答的相关变化。 此外，我们将评估这些T细胞的致脑炎活性， 通过被动转移到MHC相容的，重建的骨髓中， SCID-Hu小鼠。 AIM 2.探讨TCR肽注射液对TCR和血压的影响 应答 随着对V-beta-5.2的关注，我们将注入重叠 肽进入MS患者与V-β-5.2偏倚的反应，BP， 确定哪些区域是免疫显性T细胞和B细胞独特表位。 在 响应者，我们将监测V-β-5.2 + T细胞的功能变化， 对BP的回应 AIM 3.评估TCR诱导的潜在调节机制 缩氨酸 为此，我们将分离并表征不同的 抗TCR肽特异性T细胞克隆型和诱导的抗体 TCR肽增强。 我们将评估每个T细胞克隆型， 亲和纯化的抗体调节自体免疫的能力 体外和SCID-Hu小鼠中的V-β-5.2 + BP反应性T细胞。