睾丸支持细胞与共刺激通路阻断剂对异体移植肾细胞的保护作用

Experimental study of sertoli cell and CTLA-4Ig′s induction of immune tolerance of allogeneic renal cell

目的 研究表达Fas配体 (FasL)的睾丸支持细胞与共刺激通路阻断剂细胞毒性T细胞相关抗原 4免疫球蛋白 (CTLA 4Ig)对异体移植肾细胞的保护作用 ,以提高移植肾的免疫耐受性。方法 酶消化法制备睾丸支持细胞及肾细胞。取 10 6个细胞混合移植于大鼠肾包膜下 ,实验大鼠分为 3组 :对照组 (10只 )、混合细胞移植组 (混合移植组 ,16只 )、联合CTLA 4Ig组 (CTLA组 ,10只 )。分别于术后 1、7、14、2 0d检测血清白细胞介素 2 (IL 2 )水平变化。术后 2 0d取出移植物 ,以亲和素 生物素 过氧化物酶复合物技术观察肾细胞存活状况 ;MD 2 0图像分析系统测定混合移植物灰度值 ;末端脱氧核糖核酸转移酶介导的X dUTP缺口末端标记 (TUNEL)法观察移植物中凋亡的细胞。 结果对照组无移植物存活 ;混合移植组 14只移植物存活 ,灰度值为 0 36 2± 0 0 17;CTLA组 10只移植物均存活 ,灰度值为 0 4 4 5± 0 0 2 1;混合移植组与CTLA组间灰度值差异有显著意义。术后CTLA组血清IL 2水平较混合移植组低 ,差异有显著意义 ;混合移植物中可见凋亡的淋巴细胞。 结论 异体肾细胞移植中 ,睾丸支持细胞与CTLA 4Ig对移植肾细胞具有协同保护作用。

Objective To study the protective role of Sertoli cell expressing FasL and CTLA 4Ig on allogeneic renal cell. Methods Testicular Sertoli cell and renal cell were prepared by digestion with enzymes. About 106 cells were injected into the subrenal capsule of allogeneic rats. 36 rats were divided into 3 groups: control group(10), co transplantation group(16) and co transplantation collaborated with CTLA 4Ig group(CTLA group,10).Levels of serum IL 2 were tested on the 1st, 7th, 14th, 20th day after transplantion. The kidney was extracted on the 20th day. The survival of renal cells was determined by the Avidin Biotin Peroxidase Complex Technique (ABC); the brightness of grafts was measured by MD 20 image analysis system. Apoptosis within the grafts was observed with Terminal Deoxynucleotidyl Transferase mediated X dUTP Nick end Labeling (TUNEL) method. Results The survival of renal cells in control group, co transplantation group and CTLA group was 0,14 and 10 respectively; The brightness values of co transplantation group and CTLA group were 0 362±0 017 and 0 445±0 021, and the statistic differences between them were significant. Levels of serum IL 2 in CTLA group were lower compared with co transplantation group, there being significant differences as well; Apoptosis of lymphocyte was observed within the grafts. Conclusions Sertoli cell and CTLA 4Ig have coordinative protective effect on allogeneic renal cell.