重组人粒细胞集落刺激因子动员对淋巴细胞功能相关抗原1／细胞间黏附分子1信号途径诱导CD4^＋T淋巴细胞活化及增殖的影响

目的：观察在异基因外周血造血干，祖细胞移植中重组人粒细胞集落刺激因子动员对淋巴功能相关抗原1/细胞间黏附分子1信号途径诱导的外周血CD4＋T淋巴细胞的影响，验证重组人粒细胞集落刺激因子动员对淋巴细胞功能相关抗原1/细胞间黏附分子1信号通路的抑制作用。 方法：选择2006—06/2007—06在解放军总医院血液科异基因外周血造血干细胞移植前进行重组人粒细胞集落刺激因子动员的10例健康供者，对治疗方案均知情同意，且得到医院伦理道德委员会批准。给予重组人粒细胞集落刺激因子10μg，（kg·d）进行动员，在动员前1天和动员后第5天取供者外周静脉血，用miniMACS磁珠分选系统分离纯化CD4＋T淋巴细胞，分别用CD3单克隆抗体OKT3＋细胞间黏附分子1、佛波酯＋离子霉素刺激活化CD4^＋T淋巴细胞，用双色荧光标记检测动员前后CD4^＋T淋巴细胞激活后活化标记CD69，CD25的表达；用四甲基偶氮唑盐法检测动员前后OKT3＋细胞间粘黏附分子1、佛波酯＋离子霉素刺激CD4^＋T淋巴细胞增殖能力变化。 结果：①纯化后CD4^＋T淋巴细胞纯度均在90％以上。②动员后OKT3＋细胞间黏附分子1组、佛波酯＋离子霉素组CD4^＋T淋巴细胞活化标记CD69和CD25的表达均明显低于动员前（P〈0．01）。②动员后OKT3＋细胞间黏附分子1组、佛波酯＋离子霉素组CD4^＋T淋巴细胞增殖率均明显低于动员前（P〈0．05）。 结论：重组人粒细胞集落刺激因子动员可能抑制了淋巴功能相关抗原1/细胞间黏附分子1协同刺激信号，从而使CD4^＋T淋巴细胞活化、增殖能力下降。

Impact of mobilization with recombinant human granulocyte colony-stimulating factor on the activation and proliferation of CD4^＋T cells through a signal way of lymphocyte functionassociated antigen-1/intracellular adhesion molecule-1

AIM： To study the impact on activation and proliferation of donor＇s CD4^＋ T cells which are induced by lymphocyte antigen-1 （LFA-1）/intraceUular adhesion molecule-1 （ICAM-1） costimulatory signal during mobilization with recombinant human granulocyte colony-stimulating factor （rhG-CSF） in the allogene graft of peripheral blood hematogenesis stem/progenitor cells, and verify the inhibitory effect of rhG-CSF on LFA-1/ICAM-1 signal.
METHODS： The experiment was performed in the Laboratory of Hematology, Chinese PLA General Hospital from June 2006 to June 2007, with the approval of the hospital ethics committee. Informed consents were obtained from all the subjects. Ten donors received rhG-CSF with the dose of 10 μg/kg per day. The peripheral blood mononuclear cells were collected one day before mobilization and on the fifth day of mobilization with rhG-CSF. And CD4^＋ T cells were purified by miniMACS. After activated by OKT3＋ICAM-1 and phorbol myristate acetate （PMA）＋ion separately, activation marker CD69 and CD25 were detected by flow cytometry, and proliferation was testing by methyl thiazolyl tetrazolium assay.
RESULTS： ①The purity of CD4^＋ T cells after activation exceeded 90%. ②In OKT3＋ICAM-1 group and PMA＋ion group, the expressions of CD69 and CD25 after mobilization were significantly lower than those before mobilization （P 〈 0.01 ）. ③The proliferation rate of CD4^＋T cells after mobilization of OKT3＋ICAM-1 and PMA＋ion was significantly lower than that before mobilization （P 〈 0.05）.
CONCLUSION： Mobilization with rhG-CSF may inhibit the activation and proliferation of CD4^＋T cells through inhibiting LFA-1/ICAM-1 costimulatory signal.