Allegretta M, Steinman L
Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Beckman Center for Molecular and Genetic Medicine (B002), California 94305, USA.
Ann N Y Acad Sci. 1995 Jul 7;756:265-82. doi: 10.1111/j.1749-6632.1995.tb44524.x.
We have used two approaches to isolate TCR sequences that are unique to patients with multiple sclerosis. One strategy was to sequence TCR gene rearrangements directly from MS lesions. The second strategy utilized T-cell clones with a selectable mutation that are found only in MS patients. The selection of T-cell clones with mutations in the hypoxanthine guanine phosphoribosyltransferase (hprt) gene was used to isolate T-cells reactive to myelin basic protein (MBP) in patients with multiple sclerosis (MS). These T-cell clones are activated in vivo, and are not found in healthy individuals. The third complementarity determining regions (CDR3) of the T-cell receptor (TCR) alpha and beta chains are the putative contact sites for peptide fragments of MBP bound in the groove of the HLA molecule. The TCR V gene usage and CDR3s of these MBP-reactive hprt- T-cell clones are homologous to TCRs from other T-cells relevant to MS, including T-cells causing experimental allergic encephalomyelitis (EAE) and T-cells found in brain lesions and in the cerebrospinal fluid (CSF) of MS patients. In vivo activated MBP-reactive T-cells in MS patients may be critical in the pathogenesis of MS.
我们采用了两种方法来分离多发性硬化症患者特有的TCR序列。一种策略是直接从MS病变中对TCR基因重排进行测序。第二种策略利用仅在MS患者中发现的具有可选择突变的T细胞克隆。通过选择次黄嘌呤鸟嘌呤磷酸核糖转移酶（hprt）基因发生突变的T细胞克隆，来分离多发性硬化症（MS）患者中对髓鞘碱性蛋白（MBP）有反应的T细胞。这些T细胞克隆在体内被激活，在健康个体中未发现。T细胞受体（TCR）α和β链的第三个互补决定区（CDR3）是与结合在HLA分子沟槽中的MBP肽片段的假定接触位点。这些对MBP有反应的hprt-T细胞克隆的TCR V基因使用情况和CDR3与其他与MS相关的T细胞的TCR同源，包括引起实验性变应性脑脊髓炎（EAE）的T细胞以及在MS患者脑病变和脑脊液（CSF）中发现的T细胞。MS患者体内被激活的对MBP有反应的T细胞可能在MS的发病机制中起关键作用。
