不同免疫抑制剂诱导外周血活化T淋巴细胞凋亡的研究  

Induction of apoptosis of peripheral activated T lymphocytes by immunosuppressants

作　　者：肖序仁[1] 徐衍盛[1] 敖建华[1] 

机构地区：[1]解放军总医院泌尿外科,北京100853

出　　处：《中华器官移植杂志》2006年第11期644-647,共4页Chinese Journal of Organ Transplantation

摘　　要：目的探讨各种免疫抑制剂在体外对人外周血淋巴细胞凋亡的诱导作用及其机制。方法从健康人外周血中分离T淋巴细胞,经抗原刺激诱导淋巴细胞活化,将霉酚酸酯(MMF)、环孢素A(CsA)、他克莫司(FK506)、西罗莫司、泼尼松(Pred)、达利珠单抗及硫唑嘌呤(Aza)单独或联合加入静止T淋巴细胞和活化T淋巴细胞中共同培养,3d后,采用共聚焦显微镜、流式细胞仪、DNA片段化电泳分析及逆转录聚合酶链反应基因扩增方法检测淋巴细胞凋亡发生率,酶联免疫吸附试验检测试剂盒测定细胞培养基中白细胞介素2(IL-2)和Fas的表达水平。结果各种免疫抑制剂中,仅Pred能明显促进静止T淋巴细胞凋亡。MMF、Aza及Pred均能促使活化T淋巴细胞发生凋亡(P＜0．05,P＜0．01),而CsA、FK506、西罗莫司及达利珠单抗均抑制活化T淋巴细胞的凋亡(P＜0．01)。两种或三种不同的免疫抑制剂同时加入活化T淋巴细胞的培养基中,其凋亡细胞明显少于对照组(P＜0．01)。FK506和CsA能显著抑制活化T淋巴细胞的Fas和IL-2表达(P＜0．05)。结论MMF、Aza及Pred可能通过Fas/FasL信号通路诱导活化T淋巴细胞的凋亡,而CsA和FK506通过抑制IL-2的产生而阻碍活化T淋巴细胞的凋亡,西罗莫司和达利珠单抗则可能通过阻断IL-2的活化效应而抑制活化T淋巴细胞的凋亡。Objective To investigate the effects and mechanisms of immunosuppressants on induction of apoptosis of peripheral T lymphocytes. Methods T lymphocytes were derived from healthy donors and activated by super antigen SEB. The rest or activated T lymphocytes were incubated with immunosuppressants such as myophenolate mofetil （MMF）, cyclosporine A （CsA）, FK506, azathioprine （Aza）, sirolimus （SRL）, prednisone （Pred）, and daclizumab （Dac, anti-CD25mAb）, alone or combined, for 3 days. The incidence of apoptosis was determined by the methods of confocal microscopy, flow eytometer, DNA ladder fragmentation eleetrophoresis, and reverse transeription-polymerase chain reaction （RT-PCR） gene amplification profiles. The quantitive assay of IL-2 and Fas in the culture medium was also performed using the enzyme linked immunosorbent assay kit. Results Apoptosis in rest T lymphocytes was just induced by Pred among various immunosuppressants. MMF, Aza, and Pred promoted apoptosis in activated T lymphocytes （P〈0. 05, P〈0. 01）, but it was blocked by CsA, FK506, SRL, and Dac （P〈0. 01）. After adding two or three kinds of immunosuppressants, the incidence of apoptosis in activated T lymphocytes was apparently lower than in control group （P〈 0. 01）. The expression of Fas and IL-2 by activated T lymphocytes was inhibited by FK506 and CsA （P〈0. 05）. Conclusion MMF, Aza, and Pred may induce apoptosis of activated T lymphoeytes via the signal pathway of Fas/Fasl. CsA and FK506 could inhibit the apoptosis of activated T lymphocytes by blocking the production of IL-2. Also, SRL and Dae can block the apoptosis of activated T lymphocyte by interfering with the effect of IL-2 on T lymphoeytes activation process.

关 键 词：细胞凋亡 免疫抑制剂 T淋巴细胞 

分 类 号：R392.4[医药卫生—免疫学]