抗CD132抗体介导同种异型抗原激活的T细胞凋亡

摘要/Abstract

摘要： 目的研究抗CD132单抗在体外对T细胞增殖的抑制作用以及其可能的临床应用价值。方法分离BALB/c、C57BL/6小鼠的脾细胞进行双向混合淋巴细胞培养(MLC)。分别于第一天(组1)或第三天(组2)加入抗CD132单抗(终浓度达到100mg·L^-1)。用流式细胞技术检测细胞增殖(CFSE), T细胞凋亡(PE-CD3, FITC-Annexin-v)以及细胞分裂周期(propidium iodide estain)。T细胞Survivin的表达则用免疫化学染色法检测。结果混合淋巴细胞培养的结果显示CFSE标记的脾细胞出现了不同程度的荧光强度减弱, 提示不同分裂次数的细胞存在。与MLC组比较, 组1和组2中均未发现有进一步细胞分裂的迹象存在。在第3天时, 组1可以检测到部分T细胞凋亡, 而在培养的最初两天并无此现象。在组2中, 第4天处于G2/M期的细胞出现减少而凋亡细胞增多。对照组无此现象发生(P<0.01)。同时, 在MLC组中可检测到survivin在T细胞表达, 而在组1和组2中则未能检测到。结论研究表明阻断CD132信号途径可以通过诱导同种异型抗原活化的T细胞凋亡而抑制T细胞增殖。因此, 抗CD132单抗很有希望成为器官移植中抗排斥的临床药物。

关键词: CD132, CD132, CD132, 凋亡, 凋亡, 凋亡, 混合淋巴细胞培养, 混合淋巴细胞培养, 混合淋巴细胞培养

Abstract: Aim To investigate the mechanism of anti-CD132 monoclonal antibodies (mAbs) inhibiting T cells proliferation in vitro, and their potential values for clinical use. Methods BALB/c and C57BL/6 mice splenocytes were harvested for two-ways mixed lymphocyte culture (MLC). Anti-CD132 mAbs (final concentration 100 mg·L^-1) were added in MLC on day 0 (group 1) or day 3 (group 2). Fluorescence activated cell sorting (FACS) was used to measure the proliferation (carboxy-fluorescein dia cetate, succinimidyl ester,CFSE), apoptosis of T cells (PE-CD3, FITC-annexin-v), and cell cycle (propidium iodide stain). The expression of survivin in T cells was detected by immunochemical stai- ning. Results Multi-generation CFSE-labeled splenocytes were found dividing and their fluorescent strength decreased in MLC. There was no noticeable change in fluorescent intensity in group 1 and group 2. On day 3, apoptosis induced by anti-CD132 mAbs was detected in part of T cells, but was not detected in the former two days in group 1. In group 2, the number of cells in M phase (activated T cells) decreased and apoptotic cells increased on day 4. The phenomena were not observed in control group (P<0.01). Expression of survivin in T cells was detected in control group but not in groups 1 and 2. Conclusion Blockade of CD132 signaling pathway inhibits T cell proliferation in vitro by means of inducing activated alloreactive T cell apoptosis but not the resting T cells. Anti-CD132 mAbs may be candidates for clinical applications.

Key words: CD132, CD132, apoptosis, apoptosis, two-ways mixed lymphocyte culture, two-ways mixed lymphocyte culture

中图分类号:

R392.4

R392.12

Supporting: Foundation item: National Basic Research Program of China (973 Program , No. 2003CB515505) National Natural Science Foundation of China (No.30271243).^*Corresponding author. Tel.: 86-27-83662892

陈必成, 昌盛, 唐莉, 张鑫, 向芙莉, 郭晖, 陈忠华^*. 抗CD132抗体介导同种异型抗原激活的T细胞凋亡[J]. .

CHEN Bi-cheng, CHANG Sheng, TANG Li, ZHANG Xin, Xiang Fu-li, GUO Hui, CHEN Zhong-hua Klaus^*. Anti-CD132 Monoclonal Antibodies Inducing T Cells Apoptosis after Alloantigen Stimulation and Its Possible Clinical Applications[J]. .

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