PEPTIDE-MEDIATED IMMUNOTHERAPY FOR AUTOIMMUNE DISEASE

我们的目标是设计T细胞受体和MHC封闭肽 可预测的特性，可用于治疗 多发性硬化症。最初我们配制了一种非免疫原性多肽 基于髓鞘碱性蛋白(MBP)的序列 与I-Au结合的亲和力比脑源性要大得多 MBP的AC1-11肽。这种多肽可以逆转EAE，甚至可以预防 如果在EAE迹象首次出现时出现后续复发 才能出现。我们将把这些发现扩展到开发MHC的抑制剂 并在Lewis大鼠体内检测MBP多肽87-99的TCR。原因 为了寻找多肽87-99的TCR和MHC结合的抑制剂， 以下是关于一组主要TCR的一些偶然发现 多发性硬化症脑部病变的重排。我们分析了T细胞受体(TCR) 基因重排直接来自多发性硬化症大脑斑块。重新排列Vbeta 5.2 在所有具有HLADR2基因的患者的脑中都检测到了基因。一个 这些MS脑斑块中常见的Vbeta T.2-Dbeta-Jbeta序列是 与对Vbeta 5.2 T细胞的VDJ区的描述相同 克隆人。这个克隆来自一名多发性硬化症患者，他是人类白细胞抗原DR2，具有细胞毒性 对于含有MBP肽89-106的靶标。推导出的氨基酸序列 这种VDJ重排，LRG，以前也在T细胞中描述过， 从EAE病变组织中克隆的MBP多肽87-99具有特异性。VDJ 具有该MBP表位特异性的序列构成了一个大的 多发性硬化皮损中TRC Vbeta 5.2N(D)N重排的部分(40%)。 具有这些VDJ序列的T细胞引起EAE的能力，以及 这种序列在脱髓鞘病变中的流行，表明T 具有这种重新排列的TCR的细胞在MS中可能是关键的。我们已经分析了 MBP多肽87-99用于确定可能与其相互作用的部位 MHC和TCR。基于这些研究，我们设计了多肽 干扰MHC或TCR结合的MBP87-99的类似物， 包括在TCR的CDR3中发现的LRG基序，通常在 多发性硬化症。其中一些TCR拮抗剂可以预防EAE。因此，我们计划 开发干扰MBP的TCR和MHC识别的MBP类似物 肽87-99。我们将测试这些对手，以确定他们是否可以 在出现最初的疾病迹象后逆转EAE。我们会 还开发了针对人MBP肽87-99的CD4+T细胞克隆和 限制为HLADRB1*1501(DR2)，并测试来自 刘易斯老鼠系统也阻止了他们的人类同行。最终我们 希望将这些结果应用于多发性硬化的临床试验。

Our goal is to design of T cell receptor and MHC blocking peptides with predictable properties that might be used therapeutically to treat multiple sclerosis. Originally we formulated a non-immunogenic peptide based on the based on the sequence of myelin basic protein (MBP) that binds to I-Au with much greater affinity relative to the encephalitogenic peptide Ac1-11 of MBP. This peptide reverses EAE, and even prevents subsequent relapses, if given at the time that signs of EAE first begin to appear. We shall extend these findings to develop inhibitors of MHC and of TCR specific for MBP peptide 87-99 in the Lewis rat. The reason for pursuing inhibitors of the TCR and of MHC-binding of peptide 87-99, follows from some serendipitous finding regarding a major set of TCR rearrangements in MS brain lesions. We analyzed T cell receptor (TCR) gene rearrangements directly from MS brain plaques. Rearrange Vbeta 5.2 genes were detected in the brains of all patients who were HLA DR2. A common Vbeta t.2-Dbeta-Jbeta sequence in these MS brain plaques was identical to that described for the VDJ region of a Vbeta 5.2 T cell clone. This clone from an MS patient, who was HLA DR2, was cytotoxic for targets with MBP peptide 89-106. The deduced amino acid sequence of this VDJ rearrangement, LRG, was also described previously in T Cells, cloned from EAE lesions, which were specific for MBP peptide 87-99. VDJ sequences with specificity for this MBP epitope constitute a large fraction (40%) of the TRC Vbeta 5.2N(D)N rearrangements in MS lesions. The capacity of T cells with these VDJ sequences to cause EAE, and the prevalence of such sequences in demyelinated lesions, indicated that T cells with this rearranged TCR, may be critical in MS. We have analyzed the MBP peptide 87-99 to determine the putative interaction sites with MHC and with TCR. Based on these studies we have designed peptide analogues of MBP 87-99 that interfere with either MHC or TCR binding, including the LRG motif found in the CDR3 of TCR commonly transcribed in MS lesions. Some of these TCR antagonists prevent EAE. We thus plan to develop MBP analogues that interfere with TCR and MHC recognition of MBP peptide 87-99. We will test these antagonists to determine if they can reverse EAE after the first signs of disease have appeared. We shall also develop CD4+ T cell clones specific for human MBP peptide 87-99 and restricted to HLA DRB1*1501(DR2), and test whether the blockers from the Lewis rat system also block their human counterparts. Ultimately we would hope to apply these results to clinical trials in MS.