LFA-1对小鼠Treg细胞功能可塑性的影响及其机制探讨

Effects of LFA-1 on plasticity of Treg in inflammation conditions in vitro

目的 :淋巴细胞功能相关抗原-1(LFA-1)对淋巴细胞的分化、迁移及功能有重要影响。体外探讨LFA-1对i Treg细胞可塑性的影响和作用机制。方法:体外诱导产生纯度较高的野生组(wild type,WT)小鼠i Treg细胞及LFA-1基因敲除组(LFA-1^(-/-)组),将以上2种细胞置于含有IL-6、IL-23、TGF-β的完全培养基中,于第8天通过流式检测IL-17、Foxp3共表达的情况,PCR检测STAT3、ROR-γt前后表达量的变化,ELISA检测细胞因子IL-10、IL-17水平。结果:野生组(wild type,WT)及LFA-1基因敲除组(LFA-1^(-/-)组)小鼠Treg细胞在炎性环境中均展现出可塑性,LFA-1^(-/-)组小鼠IL-17/Foxp3共表达高于WT组小鼠。real-time PCR显示与WT组相比,LFA-1^(-/-)组小鼠ROR-γt、STAT3表达高于WT组(P<0.001)。与空白对照组相比,炎性环境中Treg细胞分泌IL-17增加,分泌IL-10减少;且这种改变在LFA-1^(-/-)组小鼠Treg细胞中更为显著(P<0.001)。结论 :LFA-1可能通过影响转录因子ROR-γt、STAT3表达水平影响Treg细胞的可塑性。

Objective： It has been widely accepted that lymphatic helper T cells in the lymph system have plasticity.Treg cells,as one of the helper T cell phenotypes,can be induced in different cytokine environment and reveal the existence of plasticity.In addition,lymphocyte function associated antigen 1（LFA-1） play an important role in the function and migration of lymphocyte differentiation.This study was aimed to investigate the effect of LFA-1 on Treg cells plasticity and the possibility mechanism in vitro.Methods：The high purity Treg cells in wild mice and LFA-1^（-/-） mice pre in vitro,and then cultured in completely medium containing IL-6,IL-23,TGF-beta.On the eighth day,the co-expressing of Foxp3 and IL-17 was measured by flow cytometer,and the expression of STAT3 and ROR gamma t was detected by PCR.The levels of IL-10 and IL-17 in supernatant were measured by ELSIA.Results：Both of the wild mice Treg cells and LFA-1^（-/-） mice Treg cells showed the plasticity in inflammatory conditions,but LFA-1^（-/-）mice Treg cells revealed a higher percentage of Foxp3/IL-17 co-expressing,and the expression of STAT3 and ROR gamma t in LFA-1^（-/-） mice Treg was also increased than that in the wild mice Treg cells（P0.001）.Furthermore,both of the two types of cells supernatants showed that IL-17 is higher with IL-10 decreasing than that of treatment,and the change is obvious in LFA-1^（-/-） mice Treg cells（P0.001）.Conclusion： LFA-1 has the effect on the plasticity of Treg by regulating the expression of STAT3 and ROR gamma t.