摘要目的 分析重组人粒细胞集落刺激因子(rhG-CSF)动员对健康供者骨髓及外周血CD34+造血干细胞及调节性T淋巴细胞(Treg细胞)的影响.方法 本研究共回顾分析75例健康供者,其中亲缘供者65例,无关供者10例.采用rhG-CSF动员造血干细胞(5～7.5μg·kg-1·d-1),动员期间每天监测血常规及外周血干细胞比例.亲缘供者第4天采集骨髓干细胞,第5天或5～6天采集外周血干细胞,无关供者第5天采集外周血干细胞.采用流式细胞仪对采集物CD34+细胞、总T淋巴细胞(CD3+)及Treg细胞(CD4+ CD25+ CD127-)进行测定.结果 经rhG-CSF动员后,骨髓采集物CD34+细胞占有核细胞比例为(0.13±0.07)％,外周血采集物CD34+细胞占有核细胞为(0.36±0.21)％,两者比较差异有统计学意义(u=7.167,P＜0.001).骨髓采集物CD34+细胞数为(53.25±33.59)/μl,外周血采集物为(874.33±540.15)/μl,两者比较差异亦有统计学意义(u=6.714,P＜0.001).骨髓采集物Treg细胞占淋巴细胞比例为0.81％ (0.40％,6.09％),外周血采集物为1.03％(0.41％,5.70％),二者比较差异无统计学意义(u=0.131,P=0.894).骨髓采集物的Treg细胞数量为(129.12±100.90)/μl,明显低于外周血采集物的(3 357.34±2 831.72)/μl(u=6.389,P＜0.001),总T淋巴细胞数量为(2 101.82±1 288.93)/μl,亦明显低于外周血采集物的(51 221.92±17 128.01)/μl(u=6.784,P＜0.001),但二者间Treg细胞与总T淋巴细胞比值的差异无统计学意义(0.069±0.048和0.067±0.048,t=0.211,P=0.833).结论 外周血采集物CD34+细胞明显高于骨髓采集物,部分解释了外周血造血干细胞移植后造血重建快,而调节性T淋巴细胞与总T淋巴细胞比值无差异,可能与二者移植后急性GVHD发生率相当有关.

abstractsObjective To investigate the influence of recombinant human granulocyte colony stimulating factor (rhG-CSF) on CD34+ hematopoietic stem cells and regulatory T cells in healthy donors.Method Seventy-five healthy donors were retrospectively analyzed in this study,including 65 related donors and 10 unrelated donors.All the donors were mobilized with rhG-CSF (5-7.5 μg·kg-1 ·d-1).Blood routine and the percentage of CD34+ stem cells in peripheral blood were monitored during mobilization.Bone marrow stem cells on day 4,and peripheral blood stem cells on day 5 and/or 6 were collected for related donors.Peripheral blood stem cells were collected on day 5 for unrelated donors.Flow cytometry was used to detect CD34+ cells,total T lymphocytes (CD3+),and regulatory T cells (CD4+ CD25 + CD127-).Result After rhG-CSF mobilization,the percentage of CD34+ stem cells was 0.13％ ± 0.07％ in bone marrow harvests,and 0.36％ ± 0.21％ in peripheral blood products (u =7.167,P＜0.001).When converted to absolute number,the CD34+ cells were (53.25 ± 33.59)/μl in bone marrow harvests,which was significantly lower than that (874.33 ± 540.15)/μl in peripheral blood products (u =6.714,P＜0.001).The percentage of regulatory T cells was 0.81％ (0.40％,6.09％) in bone marrow harvests,and 1.03％ (0.41％,5.70％) in peripheral blood products,without significant difference (u =0.131,P =0.894).The absolute number of regulatory T cells was obviously lower in bone marrow harvests (129.12 ± 100.90)/μl than in peripheral blood products (3357.34 ± 2831.72)/μl (u =6.389,P＜0.001).Although the absolute number of T cells was higher in peripheral blood products (51 221.92 ± 17 128.01)/μl than in bone marrow harvests (2 101.82 ± 1 288.93)/μl (u =6.784,P＜0.001),no significant difference was detected for regulatory T cells/total T cells ratio (0.069 ± 0.048 versus 0.067 ± 0.048/μl;t =0.211,P =0.833).Conclusion High frequency and absolute number of CD34+ stem cells in peripheral blood harvests may be partially associated with rapid hematopoietic reconstitution after transplantation.Similar regulatory T cells/total T cells ratio may contribute to the similar occurrence of acute graft versus host disease.