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本研究观察12例健康供者在使用10μg/(kg·day)rhGCSF动员前后白细胞总数变化。应用外周血涂片瑞氏染色对白细胞进行形态学分类,使用流式细胞术分析动员前后外周血单个核细胞中T细胞、B细胞、NK细胞和单核细胞比例的变化。结果发现,动员前1天外周血白细胞计数中位数为6.25(4.7-7.8)×109/L,其中淋巴细胞中位数为2.07(1.63-3.1)×109/L,单核细胞中位数为0.163(0.078-0.414)×109/L;动员第5天外周血白细胞计数中位数为37.47(24-72.57)×109/L,其中淋巴细胞中位数为3.22(1.46-5.31)×109/L,单核细胞中位数为1.2(0.706-3.627)×109/L。供者外周血白细胞的增加为动员前的6.26±2.14倍(P<0.01),其中淋巴细胞的增加为动员前的1.45±0.76倍(P<0.05),单核细胞数增加为动员前的7.48±4.41倍(P<0.01)。流式细胞术分析发现,动员前CD3+T淋巴细胞占外周血单个核细胞(PBMNC)比例的中位数为46.96%[(32.36-57.45)%],动员后为40.94%[(25.31-48.9)%];动员前CD4+/CD8+淋巴细胞比例为1.27±0.46,动员后为1.36±0.51;动员前CD4+CD8+T淋巴细胞占PBMNC比例的中位数为0.41%[(0.16-1.51)%],动员后为0.49%[(0.09-2.0)%];动员前CD16+CD56+NK细胞占PBMNC比例的中位数为13.98%[(4.08-25.08)%],动员后为16.65%[(12.06-33.05)%];动员前CD3+CD16+CD56+NK-T细胞占PBMNC比例的中位数为2.75%[(0.37-6.38)%],动员后为3.13%[(0.46-5.95)%];动员前CD20+B淋巴细胞占PBMNC比例的中位数为9.28%[(5.97-16.33)%],动员后为9.94%[(7.36-20.41)%];动员前CD14+单核细胞占PBMNC比例的中位数为12.48%[(3.54-19.35)%],动员后为29.52%[(16.51-36.76)%]。动员后CD14+单核细胞在PBMNC中的比例比动员前增加2.87±1.51倍(P<0.05);动员前后T淋巴细胞、NK细胞、NK-T细胞、B淋巴细胞在PBMNC中的比例以及动员前后CD4+/CD8+淋巴细胞比均无显著变化(P>0.10)。结论:rhGCSF动员引起的单核细胞增加可能在异基因外周血造血干/祖细胞移植的相关事件中发挥着重要作用。
This study was to observe the changes of total white blood cells in 12 healthy donors before and after mobilization with 10 μg / (kg · day) rhGCSF. Peripheral blood smears Wright staining were used to classify the leucocytes, and the changes of the ratio of T cells, B cells, NK cells and monocytes in peripheral blood mononuclear cells before and after mobilization were analyzed by flow cytometry. The median number of peripheral blood leukocytes was 6.25 (4.7-7.8) × 109 / L on day 1 before mobilization, with a median lymphocyte count of 2.07 (1.63-3.1) × 109 / L and a median monocyte count Was 0.163 (0.078-0.414) × 109 / L. On the fifth day after mobilization, the median number of peripheral blood leukocytes was 37.47 (24-72.57) × 109 / L, and the median lymphocyte count was 3.22 (1.46-5.31) × 109 / L, the median monocyte was 1.2 (0.706-3.627) × 109 / L. The increase of peripheral leukocytes in donor was 6.26 ± 2.14 times (P <0.01) before mobilization, in which the increase of lymphocytes was 1.45 ± 0.76 times before mobilization (P <0.05), the number of monocytes increased to 7.48 before mobilization ± 4.41-fold (P <0.01). Flow Cytometry analysis found that the median pre-mobilization CD3 + T lymphocyte count was 46.96% [(32.36-57.45)%] of peripheral blood mononuclear cells (PBMNC) and 40.94% after mobilization [(25.31-48.9 )%]. The proportion of CD4 + / CD8 + lymphocytes before mobilization was 1.27 ± 0.46 and 1.36 ± 0.51 after mobilization. The median percentage of PBMNC CD4 + CD8 + T lymphocytes before mobilization was 0.41% [(0.16-1.51)%] ] And 0.49% [(0.09-2.0)%] after mobilization. The median of proportion of CD16 + CD56 + NK cells to PBMNC was 13.98% [(4.08-25.08)%] before mobilization and 16.65% 12.06-33.05)%]. Before mobilization, the median of CD3 + CD16 + CD56 + NK-T cells in PBMNC was 2.75% [(0.37-6.38)%] and was 3.13% [(0.46-5.95)% ]. The median of proportion of CD20 + B lymphocytes to PBMNC was 9.28% [(5.97-16.33)%] before mobilization and 9.94% [(7.36-20.41)%] after mobilization. Before mobilization, CD14 + monocytes accounted for PBMNC The median proportion was 12.48% [(3.54-19.35)%], and 29.52% [(16.51-36.76)%] after mobilization. After mobilization, the proportion of CD14 + monocytes in PBMNC increased 2.87 ± 1.51 times (P <0.05) before mobilization; the proportions of T lymphocytes, NK cells, NK-T cells and B lymphocytes in PBMNC before and after mobilization and before and after mobilization CD4 + / CD8 + lymphocyte ratio had no significant change (P> 0.10). Conclusion: The increase of monocytes induced by mobilization of rhGCSF may play an important role in the related events of allogeneic peripheral blood hematopoietic stem / progenitor cell transplantation.