异基因造血干细胞移植健康供者外周血干细胞二次采集时机探讨

Determination of the optimal time for second allogeneic peripheral blood stem cell harvest from healthy donors

目的探讨健康供者体内应用rhG—CSF初次采集造血干细胞以后二次外周血造血干细胞采集的时机。方法38例二次采集外周血干细胞的健康供者皮下注射rhG—CSF5μg·kg^-1·d^-1，连用5d，在第5、6天采集外周血移植物（A组）；A组供者初次采集时的资料作为对照（B组）；A组供者依据初次和二次采集的75％间隔时间分为C组（≤9个月，n=30）和D组（〉9个月，n=8）。应用流式细胞术检测各组供者外周血采集物中的淋巴细胞，CD3^＋、CD3^＋CD4^＋、CD3^＋CD8^＋、CD14^＋、CD34^＋细胞以及CD3^＋CD4^-CD8^-T细胞的数量。结果A组供者外周血采集物中淋巴细胞CD3^＋CD8^＋（25．51×10^8）和CD34^＋细胞（0．51×10^8）的中位含量显著低于B组（31．55×10^8和0．70×10^8，P〈0．05）；C组供者CD3^＋CD8^＋（23．42×10^8）和CD34^＋细胞（0．42×10^8）的中位含量显著低于B组（P〈0．05）；D组供者淋巴细胞，CD3^＋、CD3^＋CD4^＋、CD3^＋CD8^＋、CDl4^＋、CD34^＋细胞以及CD3^＋CD4CD8^-T细胞的中位含量与B组的差异无统计学意义（P〉0．05）。A、C和D三组采集物中CD4^＋与CD8^＋细胞的比值、单核细胞与CD3^＋细胞的比值以及CD3^＋C1MCD8^-T细胞与CD3^＋细胞的比值与B组的差异均无统计学意义（P〉0．05）。38名健康供者初次和二次采集的间隔时间与二次采集物中的CD34^＋细胞存在显著的相关性（r=0．357，P=0．028）。结论健康供者二次采集外周血造血干细胞的时机以初次采集的9个月后为宜，9个月内采集应适当增加循环血量以保证移植物中的免疫和造血组分能满足临床需要。

Objective To investigate the optimal time for second allogeneic peripheral blood stem cell grafts （PBSC） harvest from healthy donors after in vivo recombinant human granuloeyte colony-stimulating factor application （rhG-CSF）. Methods Thirty-eight healthy donors of second collection （group A ） were treated with subcutaneous rhG-CSFμg·kg^-1·d^-1 for five consecutive days and followed by leukapheresis on day 5 and 6. The control group （group B） was thirty-eight healthy donors who had received a first PBSC collection previously. Group A was reclassified as group C （ ≤9 months ） and group D （ 〉 9 months） according to the 75% quantile of interim time between first and second collection. The quantities of lymphocytes ofCD3^＋、CD3^＋CD4^＋、CD3^＋CD8^＋、CD14^＋、CD34^＋ cells and CD3^＋CD4^-CD8^-Tcells were determined by multi-color flow cytometry. Results The median number of CD3^＋CD8^＋（25.51×10^8）and CD3^＋ cells （0.51 ×10^8 ） in group A were significantly lower than that （31.55 ×10^8 and 0.70 ×10^8 respectively） in group B （P 〈0.05）, and so did theCD3^＋CD8^＋（23.42 ×10^8 ） and CD34^＋cells （0.42 ×10^8 ） in group C than that in group B （ P 〈 0.05 ）. There was no statistical difference in median numbers of T cell subsets, monocytes, and CD34^＋ cells between group B and group D （P 〉 0.05 ）. The cell ratios of CD4^＋ / CD8^＋ , CD14^＋/CD3^＋ and CD3 ＋ CD4^- CD8^- T/CD3 ＋ in PBSC in group A, group C, and group D were similar to that in group B （ P 〉 0.05 ）. Sperman analysis showed a positive correlation between the total CD34^＋ cells in second collection and the interval time from first to second collection （ r = 0. 357, P = 0. 028 ）. Conclusion Nine months after the first collection maybe an optimal time for the second PBSC collection. For those who undergo second PBSC collection within 9 months, more circulation blood should be extracted to ensure enough inmmunological and hematopoietic compositions.