抗CD3单克隆抗体对人CD4^＋CD25^＋T淋巴细胞凋亡和自噬的影响

目的观察抗CD3单克隆抗体对分离培养的人外周血CD4^＋CD25^＋T淋巴细胞自噬、凋亡及其分泌的代表性因子转化生长因子β（TGF-β）的影响。方法采用密度梯度离心法及尼龙棉柱法分离32例健康者外周血T淋巴细胞,磁性细胞分离器（MACS）分离得到CD4^＋CD25^＋T淋巴细胞,分别利用电镜及流式细胞仪观察、检测各组（抗CD3单克隆抗体1mg/L组、IgG1同型抗体对照组）干预72h后细胞的凋亡率、自噬率,用ELISA法检测细胞培养上清液中细胞因子TGF-β的水平。结果抗CD3单克隆抗体组CD4^＋CD25^＋T淋巴细胞自噬率、凋亡率及TGF-β水平均增加（均P〈0.01）,凋亡率和自噬率之间无相关性（P〉0.05）。结论抗CD3单克隆抗体可促进CD4^＋CD25^＋T淋巴细胞凋亡和自噬及TGF-β分泌,且自噬与凋亡间相互独立。

Apoptosis and autophagy induced by anti-CD3 monoclonal antibody in human peripheral blood CD4+CD25+T lymphocytes

Objective To observe the autophagy,apoptosis rate and TGF-β secretion in human peripheral blood CD4+CD25+T lymphocytes induced by anti-CD3 monoclonal antibody.Methods Peripheral blood T lymphocyte of 32 healthy adults were separated by Percoll (1.073 g/ml) and harvested by using nylon column.Then,CD4+CD25+T lymphocytes were isolated by (magnetic cell sorting MACS).The cultured cells were divided into different groups (anti-CD3 mAb group 1 mg/L,IgG1 isotype control group).The apoptosis and autophagy rate of the cells cultured for 72 h were analyzed by electron microscope and flow cytometry.The cell culture supernatants were collected after 72 h and analyzed for levels of TGF-β by ELISA.Results The autophagy,apoptosis rate and TGF-β secretion in human peripheral CD4+CD25+T lymphocytes of the health group were increased by anti-CD3 mAb(all P<0.01);There was no significant statistic difference in correlation between apoptosis and autophagy of T lmyphocytes.Conclusions Anti-CD3 mAb could increase the autophagy rate,apoptosis rate and TGF-β secretion of peripheral blood CD4+CD25+Tregs,and there was no significant statistic difference between autophagy and apoptosis of CD4+CD25+Tregs.