基因扫描分析肾移植前后外周血T淋巴细胞谱系的变化  

Genescan analysis of expression and clonality of TCR Vβ repertoire T cell in renal transplant patients in early stage

作　　者：马俊杰[1] 于立新[1] 李阳秋[2] 杨力键[2] 刘启发[3] 徐兵[3] 

机构地区：[1]第一军医大学附属南方医院器官移植科,广州510515 [2]暨南大学医学院血液病研究所 [3]第一军医大学附属南方医院血液病科,广州510515

出　　处：《中华器官移植杂志》2004年第1期16-19,共4页Chinese Journal of Organ Transplantation

摘　　要：目的　了解肾移植患者移植前后外周血单核细胞T淋巴细胞受体 (TCR)Vβ基因谱系的变化和是否存在T淋巴细胞的特征性克隆增殖。方法　通过逆转录聚合酶链反应 (TR PCR)扩增11例首次肾移植患者术前及术后外周血单核细胞TCRVβ2 4个亚家族基因 ,了解其表达情况 ;阳性的PCR产物进一步经荧光素标记和基因扫描的方法分析TCR的互补决定区 3(CDR3)的长度及序列。结果　 11例患者术前分别表达 1～ 10个TCRVβ亚家族 ,大部分为多克隆 ;术后第 2 0d分别表达 1～ 5个亚家族 ,绝大部分为寡克隆 ;大部分TCRVβ亚家族与术前相同。发生急性排斥反应的 1例在排斥反应冲击治疗前无TCRVβ亚家族表达 ,冲击治疗过程中表达 5个Vβ亚家族 ,排斥逆转后表达 2个Vβ亚家族 (多克隆 ) ;1例肾功能恢复延迟者在恢复中表达 4个Vβ亚家族 ,恢复后表达 5个Vβ亚家族 ,以多克隆为主。 结论　肾移植患者术前外周血T淋巴细胞存在着TCRVβ亚家族倾斜分布的现象 ,术后 2 0d倾斜分布更明显 ;急性排斥反应与某些特异性T淋巴细胞克隆有关 ,随着排斥反应的逆转 ,这些克隆随之消失 ;排斥反应逆转和肾功能恢复正常后的多克隆表达可能提示免疫状态趋于正常化。Objective To investigate the expression and clonality of TCR Vβ subfamily T cell in 11 patients undergoing chronic hemodialysis before and after renal transplantation.Methods The CDR3 of TCR Vβ 24 subfamily genes were amplified in samples of peripheral blood mononuclear cells which were drawn before hemodialysis and at 20th day post-operatively. To observe the usage of TCR Vβ repertoire, the RT-PCR products were further labeled with fluorescent and analyzed by genescan technique for CDR3 size.Results (1) One patient was attacked by acute cellular rejection (ACR) and one suffered from delayed graft function (DGF) diagnosed by renal transplant biopsy. (2) 1-10 TCR Vβ subfamilies T cells could be identified before transplantation, and most of TCR Vβ subfamily T cells expressed as polyclonality. The most frequent expression of TCR Vβ genes was Vβ3. (3) Only 1-5 TCR Vβ subfamilies T cells could be detected post-operatively, most of them expressed as oligoclonality. The TCR Vβ3 subfamilies still were the most frequently expressed (in 9 cases). (4) There were no TCR Vβ subfamilies T cells before pulse of methylprednisolone, and were 5 subfamilies during pulse of methylprednisolone expressed as oligoclonality or biclonality, 2 subfamilies after ACR expressed as polyclonality. In DGF patient, there were 4 TCR Vβ subfamilies during DGF, and 5 following DGF.Conclusion (1) The significantly skew distribution of TCR Vβ subfamily T cells could be found in all patients with chronic renal failure and chronic hemodialysis. (2) Furthermore, the skewing distributions of TCR Vβ genes came to more significant at 20th day post-transplantation than before. (3) ACR might be closely associated with some special clones of TCR Vβ subfamily T cells, which subsided immediately after ACR. (4) The expression as polyclonality after ACR and DGF may indicate that the immune function partially was inclined to normalization even though immunosuppressed.

关 键 词：肾移植 T淋巴细胞 T细胞受体 基因表达 外周血 外周血单核细胞 排斥反应 

分 类 号：R699.2[医药卫生—泌尿科学] R392.4[医药卫生—外科学]