抗CD3单克隆抗体对支气管哮喘患者CD4＋CD25＋T细胞凋亡和自噬的影响

目的探讨抗CD3单克隆抗体对分离培养的支气管哮喘（简称哮喘）患者外周血CD4＋CD25＋T细胞凋亡和自噬及其分泌的代表性因子转化生长因子p（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例哮喘患者（哮喘组）及30名健康者（对照组）外周血T细胞，磁性细胞分离器分离得到CD4＋CD25＋T细胞，分别利用电镜及流式细胞仪观察、检测抗CD3单克隆抗体干预72h的细胞凋亡率、自噬率。用EI，ISA法检测细胞培养上清液中细胞因子TGF-β的水平。结果抗CD3单克隆抗体干预72h后两组外周血CD4＋CD25＋T细胞凋亡率、自噬率及TGF-β均增加（P值均〈0．01），但哮喘组凋亡率、自噬率均低于健康对照组（P值均〈0．01）；两组间 TGF-β水平无显著差异（P〉0．01）。哮喘组外周虹CD4＋CD25＋T调节细胞在CD3单克隆抗体的干预下自噬与TGF-β的分泌呈显著负相关（r=-0．38，P〈0．01）。结论抗CD3单克隆抗体可促进CD4＋CD25＋T细胞凋亡和自噬及TGF-β分泌。

Apoptosis and autophagy induced by anti-CD3 monoclonal antibody in bronchial asthma patients' peripheral blood CD4+CD25+T lymphocytes

Objective To observe the effects in autophagy, apoptosis rate and TGF-β secretion of bronchial asthma （asthma） patients peripheral blood CD4＋ CD25＋ T lymphocytes induced by anti-CD3 monoclonal antibody. Methods Peripheral blood T lymphocyte of 30 healthy adults（contorl group）and 32 asthma patients （asthma group） were separated by Percoll （1.073 g/ml） and harvested by using nylon column. Then,CD4＋ CD25＋ T lymphocytes were isolated by MACS（magnetic cell sorting）. The autophagy, apoptosis,rate of the cells induced by anti-CD3 monoclonal antibody for 72 h were analyzed by electron microscope and flow eytometry. The cell culture supernatants were collected after 72 hours and analyzed for levels of TGFq3 by ELISA. Results The autophagy and apoptosis rate of peripheral CD4＋ CD25＋ T lymphoeytes of both the health group and the asthma group were increased by CD3 mAb,but the effect was less in the asthma group than the health group（ P 〈0.01）. There was no obvious change of TGF-β among two groups（ P〉 0.05）. There was a negatively correlation between the autophagy expression and TGF-β secretion of asthmatic patients peripheral blood CD4＋CD25＋ Tregs （ r = -0.38, P d0.05）. Conclusions Anti-CD3 mAb could increase the apoptosis rate, autophagy rate and of TGF-β3 secretion of asthmatic patients peripheral blood CD4＋CD25 ＋ Tregs.