LFA-1对小鼠Treg细胞功能可塑性的影响及其机制探讨
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国家自然科学基金(81170359)


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

    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iTreg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iTreg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-γ t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-γ t 、STAT3表达水平影响Treg细胞的可塑性。

    Abstract:

    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(P<0.001). 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(P<0.001) . Conclusion: LFA-1 has the effect on the plasticity of Treg by regulating the expression of STAT3 and ROR gamma t.

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夏 菲,李 芳,张东波,于成功. LFA-1对小鼠Treg细胞功能可塑性的影响及其机制探讨[J].南京医科大学学报(自然科学版),2017,(12):1543-1547

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  • 收稿日期:2017-01-03
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  • 在线发布日期: 2017-12-25
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