目的观察Th22细胞在小鼠外周神经异种移植排斥反应中的动态变化。方法40只C57小鼠按随机数字表法分为对照组和实验组,每组20只。实验组小鼠行异种异体坐骨神经移植(供体为预先切除的SD大鼠坐骨神经),对照组小鼠行自体坐骨神经移植。利用Catwalk系统检测2组小鼠术后坐骨神经功能的变化情况:应用免疫组化染色检测移植神经周围组织中IL-22阳性细胞不同时间点的浸润情况。流式细胞术检测脾细胞Th22细胞比例的变化:ELISA法检测血清IL-22的水平。结果(1)Catwalk系统检测结果显示,实验组小鼠主要指标均明显差于对照组小鼠,差异均有统计学意义(P〈0.05)。(2)免疫组化染色结果显示,实验组小鼠移植神经周围组织中浸润的IL-22阳性细胞数量呈现出逐渐增多的趋势,并且在术后第5、15、25天显著高于对照组小鼠,差异均有统计学意义(P〈0.05)。(3)流式细胞仪检测结果显示,实验组小鼠脾内T1122细胞百分比在术后第5、15、25天分别为(7.76±0.81)%、(9.6±0.73)%、(8.08±0.66)%,明显高于对照组的(5.10±0.28)%、(5.13±0.35)%、(4.92±0.34)%,差异均有统计学意义(P〈0.05)。(4)ELISA结果显示,实验组小鼠术后第5、15、25天血清IL-22水平分别为(8.99±0.94)pg/mL、(11.0±1.08)pg/mL、(8.65±1.04)pg/mL,明显高于对照组的(5.16±0.13)pg/mL、(5.22±0.22)pg/mL、(4.96±0.29)pg/mL),差异均有统计学意义(P〈0.05)。结论Th22细胞可能参与小鼠外周神经异种移植排斥反应发生过程。
Objective To evaluate the dynamic changes of Th22 cells in rejection of the peripheral nerve xenotransplantation in mice. Methods Forty healthy male C57 BL/6 mice were randomly divided into isograft group and xenograft group (n=20). Catwalk automated gait analysis system was used to evaluate the motor function changes after isograft/xenograft treatment; immunohistochemical staining was used to observe to the interleukin (IL)-22 positive cell invasion degrees around the grafts at different time points. The percentages of Th22 cells in spleen were detected by flow cytometry; serum IL-22 levels were assessed by ELISA. Results Catwalk-assisted gait analysis showed that xenograft group had significantly poorer motor and neurological functions as compared with isograft group (P〈0.05). The degrees of IL-22^± cell invasion around the grafts in xenograft group significantly increased 5, 15, and 25 d after xenotransplantation as compared with those in the isograft group (P〈0.05). As compared with those in the isograi± group ([5.10±0.28] %, [5.13±0.35] % and [4.92±0.34] %), markedly increased percentages of Th22 cells ([7.76±0.81] %, [9.6±0.73] % and [8.08±0.66] %) were noted in the xenograft group 5, 15, and 25 d aider xenotransplantation (P〈0.05). ELISA results revealed that the serum IL-22 levels during rejection in the xenograft recipients ([8.99±0.94] pg/mL, [11.0±1.08] pg/mL and [8.65 ±1.04] pg/mL) were statistically higher than those in the control group ([5.16±0.13] pg/mL, [5.22±0.22] pg/mL and [4.96±0.29] pg/mL) 5, 15, and 25 d after xenotransplantation (P〈0.05). Conelnsion Th22 cells contribute to the rejection process of peripheral nerve xenotransplant.