抗γ公共链抗体抑制T淋巴细胞增殖的试验  

Anti-gamma common chain (γc) antibody induces T cells apoptosis after being activated by alloantigen

作　　者：陈必成[1] 昌盛[1] 唐莉[1] 张鑫[1] 郭晖[1] 王雪琼[1] 陈忠华[1] 

机构地区：[1]华中科技大学同济医学院同济医院器官移植研究所

出　　处：《中华微生物学和免疫学杂志》2005年第8期638-641,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金项目(30271243)

摘　　要：目的探讨抗γ公共链抗体体外抑制T细胞增殖的机理。方法分离BALBc、C57BL6脾细胞进行双向混合淋巴细胞培养(MLC)。第1组于MLC同时加入抗γ公共链抗体;第2组于MLC第3天加入抗γ公共链抗体。MLC为阳性对照,单独BALBc小鼠脾细胞培养为阴性对照。1～5d取培养细胞,检测培养细胞的增殖(羧基荧光素乙酰乙酸CFSE染色)、细胞凋亡(PECD3,FITCannexinⅤ)、细胞周期(PI)。脾细胞survivin表达通过免疫细胞染色方法检测。结果阳性对照在第3天开始检测到明显的分裂峰而第1、2组都未出现分裂峰。第1、2组在第1～2天未发现细胞凋亡增加,分别于第3、4天检测到部分T细胞凋亡、M期细胞数下降及凋亡细胞数增加;第2组与阳性对照组M期细胞的百分比例差异有统计学意义[(25.97±12.85)%vs(6.36±4.81)%,P=0.008)]。MLC第3天后,在阳性对照组检测到survivin表达;阴性组及试验组均未检测到survivin,其中阳性对照组与第1组的survivin阳性细胞数百分比差异有统计学意义[(18.00±14.13)%vs(1.57±1.61)%,P=0.043]。结论阻断γ公共链可以通过诱导同种反应性T细胞凋亡途径从而抑制T细胞增殖。Objective To investigate the mechanism of anti-γc McAbs inhibiting T cells proliferation in vitro. Methods BALB/c and C57BL/6 splenocytes were harvested for two way mixed lymphocytes reaction（MLC, control group）. Anti-γc McAbs （100 mg/L） were added in MLR on day 0 （group 1） and day 3（group 2）. Fluorescence activated cell sorting was used to measure the proliferation （CFSE）, apoptosis of T cells（PE-CD3, FITC-annexin-Ⅴ ） and cell cycle（PI）. The expression of survivin in splenocytes was detected by immunocytochemistry. Results CFSE labeled splenocytes were found division and fluorescent strength decreased in positive control group. There was no noticeable change in division or fluorescent intensity in group 1 and group 2. On day 3, antiγc McAbs induced apoptosis was detected in partial T cells, but not in first 2 days. In group 2, the number of cells in M phase（ activated T cells） decreased and apoptotic cells increased after 4 days MLC. The phenomena weren＇t observed in control group [（25.97 ±12.85）% vs （6.36±4.81）%, P=0.008）]. Expression of survivin in T cells was detected in control group but not in group 1 [（18.00± 14.13）% vs （1.57±1.61）%, P=0.043）]. Conclusion Blockade of γc signaling can inhibit T cells proliferation in vitro through inducing activated alloreacfive T cells apoptosis but not in resting T cells.

关 键 词：γ公共链 凋亡 SURVIVIN T细胞 T淋巴细胞增殖 SURVIVIN表达 抗体 试验 混合淋巴细胞培养 BALB/C小鼠 

分 类 号：R392[医药卫生—免疫学]