自体外周血造血干细胞移植过程中淋巴细胞亚群和CD34^+细胞变化的动态研究

Kinetic study on the variety of lymphocytes and CD34^+ cell subsets during autologous peripheral blood stem cell transplantation

目的动态观察动员、采集过程中外周血及造血干细胞中的淋巴细胞亚群和造血干细胞含量变化,及时指导临床选择最佳采集时机。方法 采用流式细胞术(FCM)测定大剂量化疗(HDC)联合重组人粒细胞集落刺激因子(rhG-CSF)对47例恶性实体瘤患者自体外周血造血干细胞移植(APBSCT)过程中,外周血和外周血造血干细胞(PBPC)中的淋巴细胞亚群CD3、CD4、CD8、NK、CD19和造血干细胞CD34+及其亚群CD34+CD38-、CD34+ Thy1+、AC133+细胞含量的变化,同时用体外集落培养的方法评价干细胞克隆形成能力。结果 动员后外周血淋巴细胞亚群CD3、CD4、CD8、NK和CD19细胞含量均低于动员前,其中CD3、CD8含量明显低于基础状态(P=0.007,P=0.016),而动员后外周血中的CD34及其亚群含量均明显高于动员前(P<0.05)。动员后中位时间第16天(15～17 d)外周血中的CD34含量达到最高峰,且第1次采集物中的造血干细胞含量最高。PBPC中除CD4、CD4/CD8明显低于外周血外(P<0.000 5),其他淋巴细胞亚群含量与外周血比较无明显改变。外周血单个核细胞中CD34+细胞含量与PBPC中CD34+细胞、CD34+CD38-及粒/单系集落形成单位(CFU-GM)、红系爆式集落形成单位(BFU—E)均存在显著相关性(P<0.05),而与采集的单个核细胞(MNC)总数、CD34+Thy1+、AC133+细胞含量间无相关关?

Objective To monitor the variety of lymphocytes and CD34+ cell subsets in peripheral blood (PB) and peripheral blood progenitor cell (PBPC) during mobilization and collection, and determine the optimal time for PBPC collection. Methods Flow cytometry (FCM) was used to detect the concentration of lymphocyte subsets, such as CDS,CD4,CDS,NK and CD19, as well as CD34+ cell subsets, such as CD34+ CD38\CD34 + Thyl + and AC133+ in PB or PBPC during autologous peripheral blood stem cell transplantation (APBSCT) during high dose chemoradiotherapy (HDC) with recombinant human granulocyte colony-stimulating factor (rhG-CSF) in 47 consecutive patients with malignant solid tumor. Colony culture in vitro was performed when evaluating the clonogenic capacity of the PBPC. Results After mobilization, the concentration of lymphocyte subsets in PB decreased to the level below the baseline with CD3 (P = 0.007) and CD8 ( P = 0.016) decreased significantly. On the contrary, the total CD34+ cell and the other CD34 subsets in PB increased significantly ( P < 0. 05). CD34+ cell peak of the blood was observed with the median time of 16 d (15-17 d) from the mobilization. The concentration of progenitor cell was the highest in the first apheresis. There was no significant change in PBPC lymphocyte subsets , except CD 4 and CD 4 / CD 8 which decreased below PB ( P < 0.000 5 ) . There was a significant linear correlation between PB CD34+ cells and PBPC CD34+ cells,CD34+ CD38\CFU-GM or BFU-E (P<0.05), but none between PB CD34 + cells and PBPC MNC,CD34+Thyl + or AC133 + cells. There was a significant correlation between PBPC CD34+ cells and PBPC CD34+ CD38\CD34+ Thyl+,MNC, CFU-GM or BFU-E (P<0.05), but none between PBPC CD34+ ceUs and AC133+ cells. After transplantation, the CD4/ CD8 ratio decreased markedly and CD19 cells returned to the normal range rapidly. Conclusion PBPC increases significantly during mobilization by HDC and rhG-CSF. The immunosuppressed state exists after APBSCT. The peripheral blood CD34 cell count is able to predict autograft yield reliably and can be the best indicator for the clinical collection of PBPC.