摘要
为了解rhG-CSF动员对供者T细胞增殖和细胞毒的影响,在rhG-CSF动员前和动员后第5天抽取供者外周血分离单个核细胞(PBMNC),在PBMNC中加入CD3单克隆抗体和rhIL-2以激活T细胞,培养96小时后检测T细胞增殖率和细胞毒性,并进一步对动员前后激活的T细胞用流式细胞术检测Fas表达率,用RT-PCR检测穿孔素(perforin)和Fas配体(FasL)mRNA表达情况,应用放射免疫分析方法检测细胞的γ干扰素(IFN-γ)分泌情况。结果发现,供者外周血T细胞增殖能力在应用rhG-CSF后下降(68.5±15.1)%(P<0.05);动员后CD3单克隆抗体和rhIL-2激活后T细胞对K562细胞的细胞毒性均比动员前细胞有显著减弱(P<0.05);动员前后T细胞在激活后的Fas表达率无明显差异(P>0.10);动员前后T细胞在激活后均表达perforinmRNA,不表达FasLmRNA;动员前T细胞激活后分泌IFN-γ的能力明显高于动员后(P<0.01)。结论:rhG-CSF动员致使供者T细胞在CD3单克隆抗体与rhIL-2激活后的增殖率和细胞毒性下降。
The study was to understand the impact on the proliferation and cytotoxicity of donor＇ s T cells during mobilization with rhG-CSF, The peripheral blood mononuclear cells （PBMNC） were collected from 15 donors before mobilization and on fifth day of mobilization with rhG-CSF, After the PBMNC were activated with 500 ng/ml of CD3 monoclonal antibody and 500 μg/ml of rhIL-2 for 96 hours, the activated T cells were collected for testing proliferation, cytotoxicity,Fas expression, perforin and Fas ligand （FasL） mRNA expression, the IFN-γ concentration in the culture medium of the activated T cells was determined by radioimmunoassay, The results showed that the proliferation activity of T lympbocytes and the cytotoxicity of T cells activated with CD3 monoclonal antibody and rhIL-2 were reduced markedly after mobilization with rhG-CSF （ P 〈 0.05 ）. The Fas molecule expression in the activated T cells was very high both before and after mobilization with rhG-CSF（ P 〉 0. 10）. The activated T cells expressed perforin mRNA and didn＇ t express FasL mRNA both before and after mobilization with rhG-CSF. The concentration of IFN-γ in the culture medium of the activated T cells decreased significantly after mobilization with rhG-CSF（ P 〈 0.01 ）. It is concluded that activity of proliferation and cytotoxicity of donors T cells is impaired after mobilization with rhG-CSF.
出处
《中国实验血液学杂志》
CAS
CSCD
2006年第5期995-998,共4页
Journal of Experimental Hematology
基金
解放军总医院十五科研计划基金重点资助项目(01YZ05)
