rhG-CSF动员对供者T细胞增殖和细胞毒的影响.docVIP

rhG-CSF动员对供者T细胞增殖和细胞毒的影响 作者：黄文荣, 王立生, 高春记, 鲁茁壮,王华, 段海峰, 达万明 【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）%（P0.05）；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（P0.05）；动员前后T细胞在激活后的Fas表达率无明显差异（P0.10）；动员前后T细胞在激活后均表达perforin mRNA，不表达FasL mRNA；动员前T细胞激活后分泌IFN-γ的能力明显高于动员后（P0.01）。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。 【关键词】 rhG-CSF动员；T细胞增殖；细胞毒性 　　Impact of Mobilization with rhG-CSF on the Proliferation and Cytotoxicity of Donor’s T Cells 　　Abstract 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 lymphocytes and the cytotoxicity of T cells activated with CD3 monoclonal antibody and rhIL-2 were reduced markedly after mobilization with rhG-CSF（P0.05）. The Fas molecule expression in the activated T cells was very high both before and after mobilization with rhG-CSF（P0.10）. 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（P0.01）. It is concluded that activity of proliferation and cytotoxicity of do