IL-12及Th1/Th2细胞因子在IL-10修饰的树突状细胞诱导大鼠角膜移植免疫耐受中的作用

Effect of IL-12 and Th1/Th2 in corneal transplantation immune tolerance induced by IL-10 modified dendritic cells in rat

目的探讨白细胞介素-12(interleukin-12,IL-12)、Th1/Th2细胞因子在IL-10修饰的树突状细胞(dendriticcell,DC)诱导大鼠角膜移植免疫耐受中的作用。方法将雄性SD受体大鼠随机分为3组,每组14只,分别为对照组(术前3d尾静脉注射PBS0.1mL)、6-DC组(术前3d尾静脉注射第6天的imDC,每只鼠0.1mL共2×106个细胞)及IL-10-DC组(术前3d尾静脉注射IL-10修饰的imDC,每只鼠0.1mL共2×106个细胞)。以雌性Wistar大鼠为供体建立穿透性角膜移植模型。术后对受体大鼠角膜植片进行临床观察,记录角膜植片存活时间;移植术后第14天检测术眼角膜中细胞因子IL-12、Th1细胞因子IL-2、IFN-γ及Th2细胞因子IL-4、IL-10mRNA的表达。结果对照组、6-DC组及IL-10-DC组角膜植片的存活时间分别为(10.63±2.00)d、(18.88±1.36)d及(24.50±2.07)d。6-DC组、IL-10-DC组与对照组比较,角膜植片存活时间明显延长(均为P<0.01);IL-10-DC组角膜植片存活时间显著长于6-DC组(P<0.01)。术后第14天,对照组角膜组织IL-12(0.80±0.00)及Th1细胞因子IL-2(1.03±0.06)、IFN-γ(0.97±0.03)的mRNA表达水平最高,而Th2细胞因子IL-4(0.13±0.00)、IL-10(0.34±0.02)的mRNA表达最低;IL-10-DC组IL-12(0.39±0.01)及Th1细胞因子IL-2(0.61±0.01)、IFN-γ(0.43±0.02)的mRNA表达水平最低,Th2细胞因子IL-4(0.19±0.00)、IL-10(0.88±0.01)的mRNA表达最高,三组比较差异具有统计学意义。结论 IL-10修饰的树突状细胞能诱导大鼠角膜移植免疫耐受;IL-12、Th1/Th2细胞因子在角膜移植免疫中起重要作用,IL-12mRNA低表达及Th1/Th2细胞因子偏离是IL-10修饰的树突状细胞诱导角膜移植免疫耐受机制之一。

Objective To study the effect of interleukin-12（IL-12） and Th1/Th2 in corneal transplantation immune tolerance induced by IL-10 modified dendritic cells（DC） in rat.Methods Male receptor SD rats were divided into control group（injected PBS 0.1 mL via tail vein at 3 days before operation）,6-DC group（injected 2×106 6-DC 0.1 mL via tail vein at 3 days before operation）,IL-10-DC group（injected 2×106 IL-10-DC 0.1 mL via tail vein at 3 days before operation）,14 cases in each group.Female Wistar rats were set as donators to establish the penetrating corneal transplantation model.The postoperative corneal grafts were clinically observed and the survival time was recorded.The expressions of IL-12,IL-2,IFN-γ,IL-4,IL-10 mRNA at postoperative 14 days were detected by RT-PCR.Results Graft survival time in control group,6-DC group and IL-10-DC group were（10.63±2.00）days,（18.88±1.36）days and（24.50±2.07）days.Compared with control group,the survival time in 6-DC group and IL-10-DC group were obviously prolonged,there were significant differences（both P0.01）,and the IL-10-DC group was obviously longer than 6-DC group,there was significant difference（P0.01）.At postoperative 14 days,in control group the expression level of IL-12 mRNA（0.80±0.00）,IL-2 mRNA（1.03±0.06）,IFN-γ mRNA（0.97±0.03） were the highest,IL-4 mRNA（0.13±0.00） and IL-10 mRNA（0.34±0.02） were the lowest;In IL-10-DC group,the expression of IL-12 mRNA（0.39±0.01）,IL-2 mRNA（0.61±0.01） and IFN-γ mRNA（0.43±0.02） were the lowest,IL-4 mRNA（0.19±0.00） and IL-10 mRNA（0.88±0.01） were the highest,there was statistical difference among three groups.Conclusion IL-10 modified DC can induce the corneal transplantation immune tolerance of rat,IL-12 and Th1/Th2 play the key role in corneal transplantation immune,the low expression of IL-12 mRNA and Th1/Th2 deviation is one of the mechanisms of transplantation immune tolerance induced by IL-10 modified DC.