Slachta C A, Jeevanandam V, Goldman B, Lin W L, Platsoucas C D
Departments of Microbiology and Immunology, Surgery, and Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, PA 19140, USA.
J Immunol. 2000 Sep 15;165(6):3469-83. doi: 10.4049/jimmunol.165.6.3469.
Chronic cardiac allograft rejection presents pathologically as graft arteriosclerosis (GA) characterized by recipient T cell and monocyte infiltration. To determine whether oligoclonal T cells are present in coronary arteries of cardiac allografts from patients with GA, we conducted sequencing analysis of beta-chain TCR transcripts from these explanted coronary arteries using the nonpalindromic adaptor-PCR. Substantial proportions of identical beta-chain TCR transcripts in three of five patients were observed, clearly demonstrating the presence of oligoclonal T cells. TCR transcripts from the arteries of two other patients were relative heterogeneous. High proportions of identical CDR3 beta-chain TCR motifs were found in each patient. GENEBANK/EMBL/SWISS PROT database comparison of all sequences revealed that these beta-chain TCR transcripts were novel. Using Vbeta-specific PCR (independent amplification), we found in patient GA03 that the TCR transcript that was clonally expanded in the left anterior descending artery after nonpalindromic adaptor-PCR was also clonally expanded in the right coronary artery of the same allograft. These results demonstrate that this TCR transcript was clonally expanded at different anatomic sides of the cardiac allograft in a systemic manner. In two patients identical beta-chain TCR transcripts that were found to be clonally expanded in the coronary arteries of their explanted cardiac allografts were also found to be clonally explanted in endomyocardial biopsies collected 17 and 21 mo earlier from each patient. The presence of oligoclonal populations of T cells in the rejected graft suggest that these T cells have undergone specific Ag-driven proliferation and clonal expansion early on within the graft and persist throughout the post-transplantation period.
慢性心脏移植排斥反应在病理上表现为以受体T细胞和单核细胞浸润为特征的移植血管硬化（GA）。为了确定GA患者心脏移植冠状动脉中是否存在寡克隆T细胞，我们使用非回文衔接子PCR对这些切除的冠状动脉的β链TCR转录本进行了测序分析。在五名患者中的三名患者中观察到相当比例的相同β链TCR转录本，清楚地证明了寡克隆T细胞的存在。另外两名患者动脉的TCR转录本相对异质性。在每名患者中都发现了高比例的相同CDR3β链TCR基序。对所有序列进行GENEBANK/EMBL/SWISS PROT数据库比较发现，这些β链TCR转录本是新的。使用Vβ特异性PCR（独立扩增），我们在患者GA03中发现，在非回文衔接子PCR后在左前降支动脉中克隆扩增的TCR转录本在同一移植心脏的右冠状动脉中也克隆扩增。这些结果表明，该TCR转录本以系统性方式在心脏移植的不同解剖部位克隆扩增。在两名患者中，发现在其切除的心脏移植冠状动脉中克隆扩增的相同β链TCR转录本，在每名患者17和21个月前采集的心肌内膜活检中也发现是克隆扩增的。排斥移植中T细胞寡克隆群体的存在表明，这些T细胞在移植早期就经历了特定抗原驱动的增殖和克隆扩增，并在整个移植后时期持续存在。
