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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。
Objective: To observe the dynamic changes of peripheral blood stem cells, immune cell subsets and cytokines after mobilization of rhesus monkey bone marrow stem cells by GM-CSF to provide reference for clinical stem cell mobilization and treatment of diseases. Methods: Healthy rhesus monkeys were injected subcutaneously with GM-CSF 8μg / (kg · d) for 5 consecutive days. Peripheral blood was collected on days 0, 2, 4, 6, 8, and 10. Blood cells were collected for counting white blood cells The proportion of CD34 +, CD133 +, CD3 +, CD4 +, CD8 + and CD56 + cells was measured by flow cytometry (FCM). The levels of TNF-α, IL-1β and IL-2 in serum were measured by enzyme-linked immunosorbent assay. Results: The numbers and proportions of WBC, neutrophil, CD34 + and CD133 + cells increased synchronously (P <0.01), and reached the peak on the 6th day after mobilization. The number of cells were 6.4, 9.1, 117 and 163.3 times of the normal level , Of which CD34 +, CD133 + returned to normal on the 8th day while WBC and neutrophils were still higher than the normal level (P <0.05). The number of CD3 +, CD4 +, CD8 + and CD56 + cells increased, and the number of cells was 4.1, 4.0, 2.9 and 4.3 times before mobilization on the 6th day, but the proportion decreased (P <0.01) 0.001), then gradually increased to the normal level and continued to the first 10 days (P <0.05). The levels of TNF-α, IL-1β and IL-2 were significantly increased within 6 days after mobilization (P <0.01), and the levels of TNF-α and IL-1β returned to normal after 8 days The increase in concentration was large and lasted at least until day 10 (P <0.01). CONCLUSION: Mobilization of cynomolgus bone marrow stem cells for 5 days continuously increases the percentage of CD34 + and CD133 + cells in peripheral blood and increases the ratio of WBC and neutrophil, increases the absolute number of CD3 +, CD4 +, CD8 + and CD56 + cells, , IL-1β, IL-2 levels increased, indicating that GM-CSF mobilization of rhesus monkey bone marrow stem cells in the immune cells and immunomodulatory factor levels improve immune function.