趋化因子CCL22 和CCL20 协同皮肤抗原诱导的调节性T 淋巴细胞对皮肤移植的影响

目的:研究小鼠皮肤移植模型中趋化因子CCL20 和CCL22 联合调节性T 细胞对移植皮肤存活时间的影响。方法:将皮肤移植小鼠分为四组,每组3 只小鼠,分别为Treg 组、Treg+ CCL20 组、Treg+ CCL22 组与对照组。其中Treg 细胞经同种异体皮肤抗原诱导。以C57BL/6 小鼠为同种异体供体,BALB/ c 小鼠为受体行皮肤移植手术。进行皮肤移植后立即于异体皮下注射Treg 细胞2伊105 个细胞/ 鼠,体积为200 滋l。以后每日于异体皮下注射趋化因子CCL20 或CCL22,共连续注射10 d,观察并记录皮肤的存活情况。Treg 细胞定植实验:首先利用磁珠分选方法分离皮肤抗原诱导Treg 细胞,并利用锝99 标记分离到的Treg 细胞;皮下注射3 h 后,处死小鼠,用GC-2016酌放射免疫计数器检测各脏器及移植皮肤的放射性活度。结果:淤经Treg 进行干预的小鼠,其异体皮肤存活时间均显著长于手术对照组(P<0.05),而且同种异体抗原诱导Treg 在趋化因子CCL20 或CCL22 存在下异体皮肤存活时间显著高于单纯使用Treg 组和对照组(P<0.001)。于注射皮肤抗原诱导的Treg 细胞后,自体和异体皮肤移植组Treg 细胞主要分布在自体和异体皮肤,分别占注射Treg 组细胞的60%和98%;加趋化因子CCL20组和CCL22 组的Treg 主要分布在肝脏。结论:趋化因子CCL20 和CCL22 能够协同促进经皮肤抗原诱导的Treg 细胞延长异体皮肤的存活时间,这种作用效果可能与趋化因子CCL20 和CCL22 趋化Treg 细胞向肝脏大量定植相关。

Effect of chemokine CCL20 and CCL22 combined with skin antigen-induced Treg on survival time of grafted skin

Objective:To study the effect of chemokines CCL20 and CCL22 combined with skin-induced Treg on survival time of grafted skin.Methods: Skin grafting mice were divided into four groups,three mice per group,namely Treg group,Treg+ CCL20 group,Treg+ CCL22 group and control group.C57BL/6 mice were used as donor and BALB/ c as acceptor,and the Treg cells were isolated from the mice induced by skin allograft.After skin grafted,CCL20 and CCL22 were subcutaneous injection every day,which lasted for 10 day.Survival time of skin in each group were observed and recorded.The Treg colonzation experiments were performed as follows.We firstly isolated Treg with Magnetic cell sorting system(MACS) and then labled them with 99 Tcm .After that we intravenously injected them into the mice.3 hours later,the mice were sacrifced and the radioactivity of organs were detected by GC-2016 radioim-munoassay counter.Results: After Treg treated the survival time of skin grafted in antigen-induced Treg group was signifiantly longer than control group,when treg were cooperated with CCL20 and CCL22,the skin grafted showed more longer survival time than Treg and control groups(P < 0.001).After injection of induced Treg,Treg in autologous and allogeneic skin grafts goups were mainly distributed in autologous and allogeneic skin,accounting for 60% and 98% respectively.When cooperated with CCL20 or CCL22,the Treg were mainly distributed in liver.Conclusion: Chemokines CCL20 and CCL22 synergistically improved the effects of skin antigen induced Treg on survival time of skin graft,which probably related with the Treg colonization into the liver .