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目的观察异品系大鼠皮肤移植排斥反应发生发展过程中外周血γδT细胞数量变化及其γ干扰素(IFN-γ)及白细胞介素4(IL-4)的分泌情况。方法建立Wistar→SD大鼠异品系皮肤移植模型,以SD大鼠自体皮肤再植为对照,于术后0、4、8d和10～13d(排斥时)取外周血用免疫荧光染色和流式细胞仪检测γδT细胞数量,并于体外扩增该细胞,用ELISA双抗夹心法测定培养上清中IFN-γ及IL-4的含量。结果异品系皮肤移植大鼠术后外周血γδT细胞数量和培养上清中IFN-γ和IL-4水平均显著升高,且显著高于自体皮肤再植同时间点对照组。结论异品系皮肤移植排斥反应早期外周血γδT细胞即可被活化,数量增多,且能同时分泌IFN-γ和IL-4,或同时存在分泌Th1和Th2类细胞因子的2个亚群,参与排斥反应的发生过程。
Objective To observe the changes of the number of γδT cells in peripheral blood and the secretion of IFN-γ and IL-4 in the skin allograft rejection after skin allograft transplantation. Methods The Wistar → SD rat model of skin allograft was established. The skin of SD rats was replanted as the control. Peripheral blood was collected at 0, 4, 8d and 10 ~ 13d after rejection by immunofluorescence staining and flow cytometry The number of γδT cells was detected by cytometry, and the cells were expanded in vitro. The contents of IFN-γ and IL-4 in the culture supernatant were determined by ELISA. Results The number of γδT cells in peripheral blood and the levels of IFN-γ and IL-4 in the supernatant of the allograft skin allografts were significantly higher than those of the autologous skin replantation group. CONCLUSION: Allogeneic skin allograft rejection can be activated and increased in number of peripheral blood γδT cells in early stage of rejection, and it can secrete IFN-γ and IL-4 at the same time. In addition, there are two subsets secreting Th1 and Th2 cytokines, The reaction occurred.