Haas R, Möhle R, Pförsich M, Fruehauf S, Witt B, Goldschmidt H, Hunstein W
Department of Internal Medicine V, University of Heidelberg, Germany.
Blood. 1995 Apr 1;85(7):1936-43.
It was the objective of the study to characterize CD34+ hematopoietic progenitor cells from peripheral blood (PB) and bone marrow (BM) in a group of 24 cancer patients. After cytotoxic chemotherapy, R-metHu granulocyte colony-stimulating factor (R-metHuG-CSF; filgrastim, 300 micrograms daily, subcutaneously) was given to shorten the time of neutropenia as well as to increase the rebound of peripheral blood progenitor cells (PBPC) for harvesting. The proportion of CD34+ cells in the leukapheresis products (LPs) was 1.4-fold greater than in BM samples that were obtained at the same day (LP: median, 1.4% v BM: median, 1.0%, P < .01). Two- and three-color immunofluorescence showed that blood-derived CD34+ cells comprised a greater proportion of a particular early progenitor cell than CD34+ cells of bone marrow. Blood-derived progenitor cells tended to have a higher mean fluorescence intensity of CD34 and expressed significantly lower levels of HLA-DR (mean fluorescence intensity of HLA-DR: 442.6 +/- 44.9 [LP] v 661.5 +/- 64.6 [BM], mean +/- SEM, P < .01). Furthermore, the blood-derived CD34+ cells comprised a 1.7-fold greater proportion of Thy-1+ cells (LP: median, 24.4% v BM: median, 14.4%, P < .001) and expressed significantly less c-kit (LP: median, 20.5% v BM: median, 31.0%, P < .01). Three-color analysis showed that high levels of Thy-1 expression were restricted to CD34+/HLA-DRdim or CD34+/HLA-DR- cells confirming the early developmental stage of this progenitor cell subset. The proportion of CD34+/CD45RA(bright) cells representing late colony-forming unit granulocyte-macrophage (CFU-GM) was smaller in LPs compared with BM (P < .05). For an examination of BM CD34+ cells before the mobilization chemotherapy, samples of 16 patients were available. The mean proportion of c-kit expressing CD34+ cells in the bone marrow during G-CSF-stimulated reconstitution decreased 1.8-fold compared with baseline values. There was no difference in the proportion of BM-derived CD34+/Thy-1+ cells and CD34+/CD45RA+ cells between steady-state hematopoiesis and G-CSF-supported recovery. Our data suggest that during G-CSF-enhanced recovery, CD34+ cells in the PB are enriched with more primitive progenitor cells to evenly replenish the BM after the chemotherapy-related cytotoxic damage.
本研究的目的是对一组24例癌症患者外周血(PB)和骨髓(BM)中的CD34+造血祖细胞进行特征分析。细胞毒性化疗后,给予R-甲硫氨酸人粒细胞集落刺激因子(R-metHuG-CSF;非格司亭,每日300微克,皮下注射)以缩短中性粒细胞减少的时间,并增加外周血祖细胞(PBPC)的反弹以进行采集。白细胞分离产物(LP)中CD34+细胞的比例比同日获得的BM样本高1.4倍(LP:中位数为1.4%,BM:中位数为1.0%,P<0.01)。双色和三色免疫荧光显示,与骨髓来源的CD34+细胞相比,血液来源的CD34+细胞在特定早期祖细胞中所占比例更大。血液来源的祖细胞CD34平均荧光强度往往更高,而HLA-DR表达水平显著更低(HLA-DR平均荧光强度:442.6±44.9[LP]对661.5±64.6[BM],平均值±标准误,P<0.01)。此外,血液来源的CD34+细胞中Thy-1+细胞的比例高1.7倍(LP:中位数为24.4%,BM:中位数为14.4%,P<0.001),c-kit表达显著更少(LP:中位数为20.5%,BM:中位数为31.0%,P<0.01)。三色分析显示,高水平的Thy-1表达仅限于CD34+/HLA-DRdim或CD34+/HLA-DR-细胞,证实了该祖细胞亚群的早期发育阶段。与BM相比,LP中代表晚期粒细胞-巨噬细胞集落形成单位(CFU-GM)的CD34+/CD45RA(bright)细胞比例更小(P<0.05)。为了检查动员化疗前的BM CD34+细胞,有16例患者的样本可用。在G-CSF刺激的造血重建过程中,骨髓中表达c-kit的CD34+细胞的平均比例与基线值相比下降了1.8倍。在稳态造血和G-CSF支持的恢复过程中,BM来源的CD34+/Thy-1+细胞和CD34+/CD45RA+细胞的比例没有差异。我们的数据表明,在G-CSF增强的恢复过程中,PB中的CD34+细胞富含更多原始祖细胞,以在化疗相关的细胞毒性损伤后均匀补充BM。