大鼠肝移植物中T-bet^＋/GATA-3^＋和ROR-γt^＋/FOXP3^＋细胞随排斥程度的动态变化及意义

The dynamic balance of T-bet^＋/GATA-3^＋ and ROR-γt^＋/FOXP3^＋ cells in rat liver allograft according to acute rejection severity

目的 检测Th1、Th2、Th17和iTreg CD4^＋T细胞亚系特征性表达的核转录因子以探究它们在大鼠肝移植急性排斥中的动态变化及意义.方法 Kamada＂二袖套＂法建立大鼠肝移植急性排斥模型,按Banff标准对排斥程度进行分级;免疫组织化学方法检测Th1、Th2、Th17和iTreg特征性表达的核转录因子,对阳性细胞计数.结果 与对照组比较,轻度排斥组、中度排斥组和重度排斥组中T-bet^＋（对照组：19.3±5.1;轻度排斥组：63.7±9.7;中度排斥组：40.9±13.6;重度排斥组：32.3±3.3）细胞数和RORγt^＋（对照组：6.5±1.4;轻度排斥组：8.3±2.3;中度排斥组：26.8±3.2;重度排斥组：39.2±12.8）细胞数显著升高（P〈0.01）;轻度排斥组中GATA-3^＋（对照组：7.1±1.3;轻度排斥组：6.4±3.1;中度排斥组：23.2±9.1;重度排斥组：42.6±14.1）细胞数无明显升高（P〉0.05）,中度排斥组中明显升高（P〉0.05）,重度排斥组显著升高（P〈0.01）;轻度排斥组FOXP3^＋（对照组：9.5±2.4;轻度排斥组：13.5±4.8;中度排斥组：24.5±4.9;重度排斥组：39.3±11.7）细胞数无明显升高（P〉0.05）,中度、重度排斥组显著升高（P〈0.01）.结论 大鼠肝移植术后急性排斥的发生与T-bet^＋、GATA-3^＋、RORγt^＋和FOXP3^＋细胞相关;在排斥早期,T-bet^＋/GATA-3^＋平衡与排斥的发生相关性比RORγt^＋/FOXP3^＋平衡更强.

Objective To evaluate the dynamic balance of Th1, Th2, Th17 and iTreg cells in the process of acute rejection （AR） , orthotopic liver transplantation （OLT） was performed from LEW rats to BN rats. Methods OLT donated by male inbred LEW rats was performed on male inbred BN rats by Kamada＇ s two-cuff technique without liver arterial anastomosis, AR severity was graded using the Banff schema ,and the immunohistological method was applied to detect the expression of T-bet^＋, GATA-3^＋, RORγt^＋, and FOXP3^＋ cells in liver allografts. Results The number of T-bet^＋ cells （Control group： 19. 3±5. 1;Mild AR group： 63. 7±9. 7;Moderate AR group： 40. 9±13. 6;Severe AR group： 32. 3±3. 3） and RORγt ^＋ （Control group： 6. 5±1. 4;Mild AR group： 6.4±3. 1;Moderate AR group： 23. 2±9. 1;Severe AR group： 42. 6±14. 1） was significantly increased in mild, moderate and severe AR groups as compared with control group （all P〈0.01）. The number of GATA-3^＋cells （Control group： 7. 1±1. 3;Mild AR group： 13. 5±4. 8;Moderate AR group： 23. 2±9. 1;Severe AR group： 42. 6± 14. 1） showed no significant difference between mild AR group and control group （P〉0. 05） , while it was significantly higher in moderate and severe AR group （P〈0. 05 and P〈0. 01）. The number of FOXP3 ^＋ cells （ Control group;9.5±2.4;Mild AR group： 13. 5±4.8;Moderate AR group： 24.5 ±4.9;Severe AR group： 39.3±11.7） had no statistically significant difference （P〉0. 05）, but it was significantly decreased in moderate and severe AR group （all P〈0.01）. The ratio of T-bet ^＋ cells/GATA-3＋cells was more significantly associated with AR than that of RORγt^＋cells/FOXP3^＋ cells in the early stage. Conclusion There is dynamic change in T-het＋,GATA-3＋,RORγt＋and FOXP3＋cells in the liver allografts.The T-bet^＋,GATA-3^＋,RORγt^＋and FOXP3^＋cells were involved in AR,and the ratio of T-bet^＋cell/sGATA-3^＋cells was more significantly associated with AR than that of RORγt^＋cells/FOXP3^＋cells in the early stage.