猕猴骨髓干细胞动员后外周血干细胞、免疫细胞亚群及细胞因子变化

The changes of stem cells,immune cells and cytokines in peripheral blood after marrow stem cell mobilization in macaqne

目的:观察GM-CSF动员猕猴骨髓干细胞后外周血干细胞、免疫细胞亚群和细胞因子含量的动态变化,为临床干细胞动员及用于治疗疾病提供参考依据。方法:健康猕猴连续5 d,皮下注射GM-CSF 8μg/(kg.d),分别于0、2、4、6、8、10 d采集外周血,血细胞分析仪计数白细胞(WBC)总数、淋巴细胞和中性粒细胞比例,流式细胞术(FCM)测定CD34+、CD133+、CD3+、CD4+、CD8+、CD56+细胞比例,酶联免疫分析法测定血清TNF-α、IL-1β、IL-2含量。结果:WBC、中性粒细胞、CD34+、CD133+细胞数量和比例均同步升高(P<0.01),到动员第6天时达到高峰,细胞数量分别为正常水平的6.4、9.1、117和163.3倍,其中CD34+、CD133+第8天时恢复正常,而WBC、中性粒细胞仍高于正常水平(P<0.05)。CD3+、CD4+、CD8+、CD56+细胞的数量增加,细胞数在第6天时分别为动员前的4.1、4.0、2.9和4.3倍,但比例下降(P<0.01),到第6天达到最低(P<0.001),随后逐渐升高至正常以上水平并持续至第10天(P<0.05)。TNF-α、IL-1β、IL-2浓度于动员后6 d内明显升高(P<0.01),其中TNF-α、IL-1β浓度至8 d恢复正常(P>0.05),IL-2浓度升高幅度较大并至少持续至第10天(P<0.01)。结论:连续5 d动员猕猴骨髓干细胞可使外周血中CD34+、CD133+细胞比率短暂升高,WBC、中性粒细胞比例持续升高,使CD3+、CD4+、CD8+、CD56+细胞绝对数增加,TNF-α、IL-1β、IL-2浓度升高,表明GM-CSF动员猕猴骨髓干细胞可在细胞和免疫调节因子水平提高免疫功能。

AIM： To explore the changes of white blood cells（WBC）, neutrophils, stem cells, immune cells and cytokines after marrow stem cell mobilization by GM-CSF in macaqne. METHODS. Macaqne were injected with GMCSF 8 μg/（kg · d） for 5 days. The ratio, number and subpopulation of WBC were observed by Blood Cell Autoanalyzer; the ratios of CDI33＋, CD34＋, CD3＋, CD4＋, CD8＋ , CD56＋ cells were identified by FCM; and levels of TNF-α, IL-1β, IL-2 were tested by ELISA on 0, 2, 4, 6, 8, 10 day after stem cell mobiliation by GM-CSF. RESULTS. The ratios and number of CD133 ＋ cells, CD34 ＋ cells, WBC and neutrophils were significantly increased （ P 〈 0. 01 ）. The number of above cells was elevated to 6.4, 9.1, 117 and 163.3 times of normal at 6 day after stem cell were mobilized, but the ratios of CD133 ＋ cells and CD34 ＋ cell were decreased to the normal level after stem cell mobilization stopped while that of WBC and neutrocytes kept increasing over 8 d. The ratios of CD3＋, CD4＋, CD8 ＋ , CD16 ＋ were decreased during 1 to 6 d, but came to the normal level at 8 d and increased at 10 d after stem cell were mobilized and the number of those cells in blood increased to 4. 1, 4.0, 2.9 and 4.3 times when compared with the normal level. The concentration of TNF-α, IL-1β, IL-2 in peripheral blood was significantly increased and IL-2 level was increased higher and longer than that of TNF-α and IL-1β. CONCLUSION： CD133＋, CD34 ＋ cells and WBC, neutrophUs in peripheral blood are greatly increased after GM-CSF mobilization, but WBC and neutrophils keep increasing longer than that of CD133 ＋ , CD34 ＋ cells. The results of the number of CD3 ＋ , CD4 ＋ , CD8＋ , CD16＋ cells and the levels of TNF-α, IL-1β, IL-2 in peripheral blood indicat that marrow stem cell mobilization by GM-CSF can also stimulate immune cell proliferation and differentiation while increasing cytokine producing.