基础医学与临床 ›› 2015, Vol. 35 ›› Issue (3): 360-365.

• 研究论文 • 上一篇    下一篇

小鼠肝移植模型中调节性T细胞的数量和表型改变

陈国锋1,蔡国军2,李国锋3,王介川3,郑树森4   

  1. 1. 浙江大学医学院附属第二医院
    2. 杭州市萧山区第四人民医院外一科
    3. 杭州市萧山区第四人民医院
    4. 浙江大学医学院附属第一医院
  • 收稿日期:2014-02-27 修回日期:2014-12-07 出版日期:2015-03-05 发布日期:2015-03-03
  • 通讯作者: 陈国锋 E-mail:cgfeng101@126.com
  • 基金资助:
    促进记忆型T细胞迁移诱导移植免疫耐受的实验研究

The quantity and phenotype change of regulatory T cell in the model of mice liver transplantation

  • Received:2014-02-27 Revised:2014-12-07 Online:2015-03-05 Published:2015-03-03

摘要: 目的 探究小鼠肝移植模型中移植肝内免疫排斥反应程度和调节性T细胞的数量及功能随时间变化的规律,为揭示机体免疫耐受的建立机制提供理论依据。方法 同基因或异基因肝移植术后3、7、14和28天留取移植肝标本(每组每个时间点n=4),HE染色进行排斥活性指数评定(Banff评分标准),抗CD3、B220和Foxp3抗体免疫组化染色鉴定移植物内T、B淋巴细胞及调节性T细胞的浸润情况,实时荧光定量PCR检测Foxp3 mRNA表达,流式细胞术检测Treg细胞表面CLAT-4的表达。结果 异基因肝移植术后2周内发生免疫排斥反应,随后逐渐好转;移植物内浸润的炎性细胞主要为T淋巴细胞,其中Foxp3 mRNA及Foxp3阳性细胞数术后显著高于同基因组(异基因15.0+1.3 vs. 同基因2.7+1.0,P<0.05)。异基因移植肝内浸润的调节性T细胞表面CTLA-4表达也显著高于同基因组(异基因53%+3% vs. 同基因13%+2%,P<0.05)。结论 异基因肝移植术后免疫排斥反应由调节性T细胞负向调节并达到免疫耐受。

关键词: 小鼠,肝移植,免疫耐受,调节性T细胞

Abstract: Objective To research the degree of immune graft rejection, the quantity and function of regulatory T cells in the model of liver transplantation after mice liver transplantation to provide theory evidence in revealing mechanisms of immune tolerance. Method Transplantation liver samples were collected at 3d, 7d, 14d, 28d after allogeneic or syngeneic mice liver transplantation (N=4 for each group at one time point). Graft rejection score was given with HE staining. T and B lymphocytes and change of regulatory T cells were identified with IHC staining.The change of Foxp3 mRNA and CLAT-4 on surface of regulatory T cells were detected by real-time PCR and IFC respectively. Results Allogeneic graft was rejected during 2w after transplantation, and the rejection receded generally after 2w. Inflammatory cells intra-graft was mainly T lymphocytes. Foxp3 positive cells and mRNA of Foxp3 in allogeneic graft group was significantly higher than these in isogeneic graft group (15.0+1.3 vs. 2.7+1.0,P<0.05). Meanwhile, expression of CLAT-4 on regulatory T cells also was higher in allogeneic graft group compared with isogeneic graft group(53%+3% vs. 13%+2%,P<0.05). Conclusion The immunity rejection after allogeneic liver transplantation achieved by regulatory T cells negative regulation and then achieved immune tolerance.

Key words: mice,liver transplantation,immune tolerance,regulatory T cells