重组人粒细胞集落刺激因子对健康供者免疫细胞的影响及其应用安全性

Effects of recombinant human granulocyte colony-stimulating factor on immunocytes in health donors and its safety of application

目的:观察重组人粒细胞集落刺激因子(rhG-CSF)在动员造血干细胞时对健康供者外周血T细胞表面活化信号(CD4+CD28+,CD8+CD28+)、活化T细胞(CD4+CD25low)和调节性T细胞(CD4+CD25high)的影响及其应用的安全性。方法:30例造血干细胞健康供者[男19,女11,中位年龄28(14～56)岁]皮下注射rhG-CSF5μg/(kg.d),连续4～6d。用药前及停药后第1天、第3天及第7天用流式细胞术检测供者外周血CD4+T细胞和CD8+T细胞表面CD28表达的相对荧光强度(RFI),以及CD4+CD25low和CD4+CD25highT细胞分别在CD4+T细胞中的百分比;并在用药前及停药后第1天、第7天及第30天进行外周血白细胞、血小板、肝功能、肾功能以及脾超声检查。结果:CD4+CD28T细胞和CD8+CD28T细胞相对荧光强度表达在用药前分别为14.91±6.10和11.10±3.74,停药后第3天分别降至11.93±5.39和8.53±3.74;差异有统计学意义(P=0.034,P=0.033)。CD4+CD25lowT细胞在CD4+T细胞中百分比,用药前为(21.4±8.87)%,停药后第3天降至(18.23±5.89)%;差异无统计学意义(P>0.05)。CD4+CD25highT细胞在CD4+T细胞中百分比,用药前为(4.16±1.62)%,停药后第1天升至(6.43±2.46)%,差异有统计学意义(P=0.000)。白细胞计数和脾超声检查用药前分别为(5.91±1.02)×109/L和(33.76±2.76)cm2,停药后第1天分别升至(34.13±8.07)×109/L和(46.85±4.53)cm2;血小板计数用药前为(228.07±73.69)×109/L,停药后第7天降至(158.27±40.69)×109/L,差异均有统计学意义(P均=0.000)。ALT和Cr用药前分别为(28.23±7.69)IU/L和(60.70±15.86)μmol/L,停药后第1天分别为(27.17±7.23)IU/L和(61.10±16.38)μmol/L,差异均无统计学意义(P均>0.05)。结论:rhG-CSF能减弱CD28的表达,降低CD4+CD25low和增加CD4+CD25high在CD4+T细胞中百分比,停药后第7天恢复至动员前水平,对健康供者肝肾功能无不利影响。

Objective ：To observe the effects of recombinant human granulocyte colony-stimulating factor （rhG-CSF） on peripheral active signal of T cell surface antigen（ CD4^＋ CD28^＋ , CD8^＋ CD28^＋ ） , activated T cells, and regulatory T cells during mobilization of hematopoietic stem cells and its safety of application in health donors. Methods： Thirty normal hematopoietic stem cells donors （19 males, 11 females, median age 28 years, range 14 -56 years） received rhG-CSF 5 I.Lg,/kg daily subcutaneously for 4 -6 days. Relative fluorescence intensity （RFI） of CD28 antigen expression on peripheral CD4^＋ cells and CD8^＋ cells, the percentage of CD4^＋ CD25^＋and CD4^＋ CD2^5hish in CD4^＋ T cells were measured with a flow cytometry before rhG-CSF administration and on day 1, day 3, and day 7 after rhG-CSF discontinuation, respectively. The peripheral WBC and PLT counts, liver and renal functions, and ultrasonic measurement of spleen were performed on the dayl, day 7, and day 30 after rhG-CSF discontinuation. Results： Relative fluorescence intensity of CD28 antigen expression on peripheral CD4^＋ cells and CD8^＋ cells was 14.91 ±6.10 and 11.10 ±3.74 before rhG-CSF administration, respectively, and decreased to 11.93 ± 5.39 and 8.53 ± 3.74 on day 3 after rhG-CSF discontinuation,respectively. The differences were statistically significant （P = 0. 034, P = 0. 033 ）. The percentage of CD4^＋ CD25^low T ceils in CD4^＋ ceils was （21. 4^＋ 8. 87）% before rhG-CSF administration, and decreased to （ 18. 23± 5. 89）% on day 3 after rhG-CSF discontinuation. The difference was no statistically significant（ P 〉 0. 05 ）. The percentage of CD4^＋ CD25^high T ceils in CD4^＋ T ceils was （4.16 ± 1.62）% before rhG-CSF administration and increased to （6.43±2.46）% on day 1 after rhG-CSF discontinuation. The difference was statistically significant （P = 0. 000）. The WBC count and ultrasonic measurement of spleen were （5.91 ± 1.02） ×10^9/ L and （33.76 ±2.76） cm^2 before rhG-CSF administration, respectively, and increased to （34. 13 ＋ 8.07） ×10^9/L and （46.85± 4. 53） cm^2 on day 1 after rhG-CSF discontinuation ; the PLT conut were （228.07 ±73.69） ×10^9/L before rhG-CSF administration, and decreased to （ 158.27± 40.69 ） ×10^9/L on day 7 after rhG-CSF discontinuation. The differences were statistically significant （all P = 0. 000） . ALT and Cr were （28.23± 7.69） IU/L and （60.70 ±15.86） μmol/L before rhG-CSF administration, respectively, and were （27.17 ±7.23） IU/L and （61.10 ± 16.38） μmol/L on day 1 after rhG-CSF discontinuation, respectively. The differences were no statistically significant （ all P 〉 0. 05 ）. Conclusion ： rhG-CSF can decrease relative fluorescence intensity of CD28 antigen expression, decrease and increase the percentage of activated T cell and regulatory T ceils in CD4^＋ T ceils, respectively. The changes return to baseline levels prior to mobilization on day 7 after rhG-CSF discontinuation. There is no untoward effects to liver and renal functions in health donors.