T cell receptor repertoire in polymyositis: clonal expansion of autoaggressive CD8+ T cells.
T cell receptor repertoire in polymyositis: clonal expansion of autoaggressive CD8+ T cells.
复制标题
DOI：
10.1084/jem.181.5.1863
复制
发表时间：
1995-05-01
期刊：
影响因子：
--
通讯作者：
Hohlfeld R
中科院分区：
文献类型：
--
作者：
Bender A;Ernst N;Iglesias A;Dornmair K;Wekerle H;Hohlfeld R
在多发性肌炎（PM）中，CD8+ T细胞受体(TCR) α / β +细胞侵入并破坏主要组织相容性复合体i类阳性肌纤维。我们采用聚合酶链反应（PCR）和双荧光免疫细胞化学方法分析了PM患者肌肉中表达的T细胞受体（TCR）库。在患者1中，反相PCR显示优先使用TCR V α 33.1, V β 13.1和V β 5.1。6个TCR V α 33.1+克隆和7个V β 13.1+克隆中有5个具有相同的核苷酸序列。相比之下，V β 5.1+ tcr更具异质性。使用TCR V α 33、V β 13或V β 5特异性引物的独立PCR方法获得了类似的结果。从非炎性对照肌中无法扩增出TCR序列。此外，在PM肌肉中发现的TCR序列在同一患者或正常对照受试者的血液中都无法检测到。免疫组织化学证实，在该患者的肌肉病变中，V β 5.1和V β 13.1的比例过高。32%的CD8+ T细胞为V β 13.1+， 16%为V β 5.1+。然而，侵袭肌纤维的CD8+ T细胞中约60%为V β 13.1+，而10%为V β 5.1+。在患者2中，50%的T细胞为V β 5.1+，与患者1一样，这些T细胞主要位于间质区。在患者3中，75%的自体侵入性T细胞被抗v β 3单抗染色。用V β 3特异性引物扩增的15个PCR克隆序列分析显示，9个（60%）序列相同。结果表明：(a) PM肌肉中表达的TCR库非常有限；(b)自体侵入性T细胞和间质性T细胞的TCR使用之间存在分离；(c)自身侵袭性T细胞克隆扩增。
In polymyositis (PM), CD8+ T cell receptor (TCR) alpha/beta + cells invade and destroy major histocompatibility complex class I-positive muscle fibers. We combined polymerase chain reaction (PCR) and double- fluorescence immunocytochemistry to analyze the T cell receptor (TCR) repertoire expressed in muscle of PM patients. In patient 1, inverse PCR revealed a preferential usage of TCR V alpha 33.1, V beta 13.1, and V beta 5.1. Six of six TCR V alpha 33.1+ clones and five of seven V beta 13.1+ clones had identical nucleotide sequences. In contrast, the V beta 5.1+ TCRs were more heterogeneous. Similar results were obtained with an independent PCR method using primers specific for TCR V alpha 33, V beta 13, or V beta 5. No TCR sequences could be amplified from noninflammatory control muscle. Furthermore, none of the TCR sequences found in PM muscle could be detected in blood from the same patient or from a normal control subject. Immunohistochemistry confirmed that V beta 5.1 and V beta 13.1 were overrepresented in the muscle lesions of this patient. 32% of all CD8+ T cells were V beta 13.1+, and 16% were V beta 5.1+. However, approximately 60% of the CD8+ T cells that invaded muscle fibers were V beta 13.1+, whereas 10% were V beta 5.1+. In patient 2, 50% of the T cells were V beta 5.1+, and as in patient 1, these T cells were mainly located in interstitial areas. In patient 3, > 75% of the autoinvasive T cells stained with an anti-V beta 3 mAb. Sequence analysis of 15 PCR clones amplified with a V beta 3-specific primer showed that 9 (60%) sequences were identical. The results suggest that (a) a strikingly limited TCR repertoire is expressed in PM muscle; (b) there is a dissociation between the TCR usage of autoinvasive and interstitial T cells; and (c) the autoinvasive T cells are clonally expanded.