McDiarmid S V, Farmer D G, Kuniyoshi J S, Robert M, Khadavi A, Shaked A, Busuttil R W
Department of Surgery, Dumont-UCLA Liver Transplant Program 90024-1752.
Transplantation. 1994 Sep 27;58(6):690-7.
Rejection continues to be a major cause of graft loss in small intestine transplantation (SIT). We have studied, by semiquantitative reverse transcriptase PCR (rtPCR), the intragraft expression of cytokines relevant to rejection in a rat model. Heterotopic SIT grafts were performed from Lewis x Brown Norway F1 donors into Lewis recipients. The isograft control was Lewis into Lewis. Five animals in each isograft and allograft group were sacrificed on POD 3, 5, 7, 8, 9, 10, 12, and 14. mRNA was isolated from portions of the terminal ileum and rtPCR performed to amplify message for interleukin-2 (IL-2), IL-2 receptor (IL-2R), interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-alpha), and interferon gamma (IFN-gamma). Semiquantitative analysis was performed using 32P radionuclide incorporation and scintillation counting. The results were expressed as percent activity compared with beta-actin. Histologic correlation with cytokine expression was made. On POD 3 after SIT there was no evidence of rejection by histology and all cytokines studied showed no difference between the isograft and the allograft. On POD 5 the first evidence of mild rejection was seen on histology and IL-6, IFN-gamma, TNF-alpha showed a significant up regulation in the allograft that persisted through POD 14. mRNA for IL-2 was not significantly upregulated until POD 7 and persisted until POD 14. IL-2R was constitutively expressed in both isograft and allograft and was not a reliable predictor of rejection. Histologic rejection was moderately severe by POD 7 and severe between POD 8 and 14 correlating with the increasing expression of IL-6, IFN-gamma, and TNF-alpha. In summary, we have shown that increasing expression of mRNA for IL-6, IFN-gamma, and TNF-alpha not only correlated with severity of rejection but that upregulation began early when histologic evidence of rejection first occurred.
排斥反应仍是小肠移植（SIT）中移植物丢失的主要原因。我们通过半定量逆转录聚合酶链反应（rtPCR）研究了大鼠模型中与排斥反应相关的细胞因子在移植物内的表达。将Lewis×Brown Norway F1供体的异位SIT移植物移植到Lewis受体中。同基因移植对照组为Lewis到Lewis。在术后第3、5、7、8、9、10、12和14天，对同基因移植组和异基因移植组的五只动物实施安乐死。从回肠末端部分分离mRNA，并进行rtPCR以扩增白细胞介素-2（IL-2）、IL-2受体（IL-2R）、白细胞介素-6（IL-6）、肿瘤坏死因子α（TNF-α）和干扰素γ（IFN-γ）的信使。使用32P放射性核素掺入和闪烁计数进行半定量分析。结果以与β-肌动蛋白相比的活性百分比表示。对细胞因子表达与组织学进行相关性分析。SIT术后第3天，组织学上没有排斥反应的证据，所有研究的细胞因子在同基因移植和异基因移植之间均无差异。术后第5天，组织学上首次出现轻度排斥反应的证据，IL-6、IFN-γ、TNF-α在异基因移植中显著上调，并持续至术后第14天。IL-2的mRNA直到术后第7天才显著上调，并持续至术后第14天。IL-2R在同基因移植和异基因移植中均组成性表达，不是排斥反应的可靠预测指标。术后第7天组织学排斥反应为中度严重，术后第8天至14天为重度，与IL-6、IFN-γ和TNF-α表达增加相关。总之，我们已经表明，IL-6、IFN-γ和TNF-α的mRNA表达增加不仅与排斥反应的严重程度相关，而且在排斥反应的组织学证据首次出现时上调就已开始。
