REGULATION OF AUTOIMMUNITY WITH T CELL RECEPTOR PEPTIDES

这项提议的总体目标是检验这样一个假设 诱导对类似天然合成多肽的免疫 人TCRV-β-5.2独特位可调节潜在致病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 小鼠证明TCR V-β-8.2 CDR2多肽可以预防和 诱导T细胞治疗实验性脑脊髓炎的临床表现 能抑制单V-β-8.2 T细胞反应的细胞和抗体 至髓鞘碱性蛋白(BP)。最近，我们发现有偏见地使用 多发性硬化症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-beta-5.2+T的人的证实 MS斑块中具有BP特异性CDR3基序的细胞。我们现在已经展示了 注射可诱导产生抗TCR特异性T细胞和抗体 TCR V-β-5.2或V-β-6.1多肽进入MS患者。就这样，舞台上 是为了测试这些T细胞和抗体是否能够调节 V-β-5.2+T细胞，包括BP特异性T细胞。成功 对BP反应性的监管将允许对其作用进行批判性评估 BP在多发性硬化症过程中的作用，并将建立一个原型 治疗其他自身免疫性疾病的方法 V基因表达受限。 目的1.评估频率、特异度、TCR谱系和 人BP特异性T细胞的致脑性。为了实现这一目标，我们将 发展快速和重复评估多发性硬化症的能力- 血液和脑脊液中BP特异性T细胞反应的相关变化。 此外，我们将评估这些T细胞的脑源性活动 通过被动移植到MHC相容，重建骨髓 SCID-HU小鼠。 目的：建立TCR多肽注射液对TCR和BP的影响 回应。随着对V-beta-5.2的关注，我们将注入重叠 多肽进入对BP有V-beta-5.2偏向反应的MS患者 确定哪些区域是免疫优势T和B细胞独特位。在……里面 应答者，我们将监测V-β-5.2+T细胞的功能变化 作为对英国石油的回应。 目的3.评估TCR诱导的潜在调控机制 多肽。在这个目标中，我们将分离和表征不同的 抗TCR多肽诱导的T细胞克隆型和抗体 TCR肽助推剂。我们将评估每个T细胞克隆类型并 亲和纯化抗体对自体免疫调节能力的研究 V-β-5.2+BP反应性T细胞在体外和SCID-Hu小鼠体内。

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.