Ho A D, Young D, Maruyama M, Corringham R E, Mason J R, Thompson P, Grenier K, Law P, Terstappen L W, Lane T
Department of Medicine, University of California, School of Medicine, San Diego, La Jolla, CA 92093-0671.
Exp Hematol. 1996 Nov;24(13):1460-8.
To define an optimal regimen for mobilizing blood-derived progenitor cells from healthy donors for allogeneic transplantation, we have studied the early and lineage-committed CD34+ subsets in the leukapheresis products after mobilization with G-CSF (10 micrograms/kg/d), GM-CSF (10 micrograms/kg/d), and the combination of G-CSF and GM-CSF (G/GM, 5 micrograms/kg/d of each). We used three color and five dimensional flow cytometry with a panel of monoclonal antibodies against CD3, CD7, CD10, CD11b, CD15, CD33, CD34, CD38, CD45, CD61, and CD71. As reference, we also analyzed CD34+ subsets in samples from umbilical cord blood (UCB) and from adult bone marrow (BM). The level of total CD34+ cells was 0.04 +/- 0.03% (mean +/- SD) in peripheral blood at baseline, and reached a maximum on day 5 or day 6 of administration of growth factors. The percentages of CD34+ cells in the leukapheresis products were 1.06 +/- 0.37% (mean +/- SD) with G-CSF mobilization, 0.35 +/- 0.24% with GM-CSF, and 0.92 +/- 0.61% with the combination of both. Among the CD34+ subsets, the percentage of cells that were CD34+/CD38- was highest in UCB (7.18 +/- 5.58%) and lowest in G-CSF mobilized peripheral blood (0.80 +/- 0.22%), whereas GM-CSF or G/GM mobilized products gave rise to intermediate levels (4.43 +/- 3.40%, 3.61 +/- 2.42%, respectively). The differences between G/GM and G-CSF, between UCB and G-CSF, or between UCB and BM are significant. The absolute numbers of CD34+/CD38- and CD34+/CD38-/HLA-DR+ subsets are also significantly higher in the G/GM mobilized products than in G-CSF products. The cloning efficiency of G/GM mobilized CD34+ cells was 2 times higher than that of G-CSF mobilized CD34+ cells, albeit the difference was statistically marginal. The profile of CD34+ subsets mobilized by the combination of G/GM approaches that found in UCB. Our data illustrate that different growth factors and regimens can preferentially mobilize different CD34+ subsets from normal donors, and that the combination of G-CSF and GM-CSF might be an optimal regimen.
为确定动员健康供体的血液来源祖细胞用于异基因移植的最佳方案，我们研究了用粒细胞集落刺激因子（G-CSF，10微克/千克/天）、粒细胞-巨噬细胞集落刺激因子（GM-CSF，10微克/千克/天）以及G-CSF与GM-CSF联合使用（G/GM，各5微克/千克/天）动员后白细胞分离产物中的早期及谱系定向CD34+亚群。我们使用三色五维流式细胞术及一组针对CD3、CD7、CD10、CD11b、CD15、CD33、CD34、CD38、CD45、CD61和CD71的单克隆抗体。作为对照，我们还分析了脐带血（UCB）和成人骨髓（BM）样本中的CD34+亚群。基线时外周血中总CD34+细胞水平为0.04±0.03%（平均值±标准差），在生长因子给药的第5天或第6天达到最高值。白细胞分离产物中CD34+细胞百分比在G-CSF动员时为1.06±0.37%（平均值±标准差），GM-CSF动员时为0.35±0.24%，两者联合动员时为0.92±0.61%。在CD34+亚群中，CD34+/CD38-细胞百分比在UCB中最高（7.18±5.58%），在G-CSF动员的外周血中最低（0.80±0.22%），而GM-CSF或G/GM动员的产物则处于中间水平（分别为4.43±3.40%，3.61±2.42%）。G/GM与G-CSF之间、UCB与G-CSF之间或UCB与BM之间的差异具有显著性。G/GM动员产物中CD34+/CD38-和CD34+/CD38-/HLA-DR+亚群的绝对数量也显著高于G-CSF产物。G/GM动员的CD34+细胞的克隆效率比G-CSF动员的CD34+细胞高2倍，尽管差异在统计学上不显著。G/GM联合方案动员的CD34+亚群谱与UCB中发现的相似。我们的数据表明，不同的生长因子和方案可优先从正常供体中动员不同的CD34+亚群，且G-CSF与GM-CSF联合使用可能是最佳方案。
