摘要
目的探讨 CD4^+CD25^+调节性 T 细胞、共刺激通路阻断剂 CD154单抗及两者联合应用在抑制大鼠肝移植急性排斥反应中的作用。方法 48例原位肝移植大鼠(DA→Lewis)分为 A 组:对照组;B 组:术前7 d 回输经 DA 大鼠脾细胞体外激活的 Lewis 大鼠 CD4^+CD25^+细胞;C 组:术后第1、2 d 腹腔注射 CD154单抗(15 mg/kg);D 组:联合应用 CD4^+CD25^+细胞和 CD154单抗。术后7 d 各组处死6只受体,观察移植肝病理变化,检测移植肝内 T 细胞亚群和细胞因子白细胞介素-2(IL-2)、IL-4、IL-10和转化生长因子β1(TGFβ1)表达情况;分离脾淋巴细胞与供体行单向混合淋巴细胞反应观察刺激指数。余大鼠观察生存情况。结果 D 组平均存活时间(52.00±10.64)d 明显长于其他各组(P<0.01);移植肝内淋巴细胞浸润数量(2.47±0.61)×10~6和 CD8^+细胞百分比(14.2±3.0)%明显低于 B、C 组(P<0.05、P<0.01),而 CD4^+CD25^+细胞比例(16.4±4.3)%高于 B、C 组(P<0.05,P<0.01)。移植物内 IL-2 mRNA 表达 A 组最高,D 组最弱;IL-4 mRNA 各组均弱表达;IL-10mRNAB、D 组高表达,A、C 组未表达;TGFβ1 mRNA B、D 组表达明显强于 A、C 组。单向混合淋巴细胞反应 D 组刺激指数最低(P<0.05)。结论 CD4^+CD25^+调节性 T 细胞和共刺激通路阻断剂 CD154单抗均能抑制大鼠肝移植急性排斥反应;联合应用 CD154单抗明显增强 CD4^+CD25^+调节性 T 细胞对急性排斥反应的抑制作用。
Objective To investigate the effect of CD4^＋ CD25 ^＋ regulatory T cell （Treg） combined with anti-CD154 mAb, a costimulatory pathway inhibitor, on acute rejection after liver transplantation. Methods CD4^＋ T cells were isolated from the spleen of a Lewis rat and labeled with CD25-PE antibody. Anti-PE micrabeads were added to collect CD4 ^＋ CD25 ^＋ T cells. Splenocytes were isolated form DA rat, treated with mitomycin C, and then co-cultured with the CD4 ^＋ CD25 ^＋ T cells of Lewis rat for 5 days for ex vivo activation. Forty-eight Lewis rats received orthotopic transplantation of the livers of DA rats, and then were randomly divided into 4 equal groups： Group A, used as control group, Group B, undergoing intravenous injection of the CD4I^＋ CD25^＋ Treg activated ex vivo by splenocytes of DA rat 7 days before the liver transplantation, Group C, undergoing intraperitoneal injection of anti-CD154 mAb twice 1 and 2 days after the liver transplantation, and Group D, undergoing injection of both CD4 ^＋ CD25 ^＋ Treg and anti- CD154. Seven days after the transplantation 6 Lewis rats in each group were harvested to observe the pathological changes of the livers and to detect the infiltrating lymphocytes, and CD4^＋ , CD8^＋ , and CD4^＋ CD25 ^＋ T cells. RT-PCR was used to detect the mRNA expression of IL-2, IL-4, IL-10, and TGFβ1 in the liver. Mixed lymphocyte reaction （MLR） was performed to evaluate the tolerance status. The remaining rats were used to observe the survival status. Results The mean survival time of Group D was 52.120 ± 10.64 days, significantly longer than those of the other three groups （P 〈 0. 05 or P 〈 0. 01 ）. Seven days after transplantation, the number of infiltrating lymphocytes in liver of Group D was significantly lower than those in the other 3 groups （ P 〈 0. 05 or P 〈 0.01 ）, whereas the proportion of CD4 ^＋ CD25 ^＋ T cells was significantly higher than those of the other 3 groups （P 〈 0.01 or P 〈 0.05）. The mRNA expression level
出处
《中华医学杂志》
CAS
CSCD
北大核心
2007年第14期942-946,共5页
National Medical Journal of China
基金
上海市卫生局科研基金(2006010)
