Nagler A, Eldor A, Naparstek E, Mumcuoglu M, Slavin S, Deutsch V R
Department of Bone Marrow Transplantation, Hadassah University Hospital, Jerusalem, Israel.
Exp Hematol. 1995 Nov;23(12):1268-74.
Protracted thrombocytopenia and bleeding remain serious complications in bone marrow transplantation (BMT). Major progress has been made in facilitating myeloid and erythroid engraftment, but little has been made in accelerating thrombopoiesis post-BMT. We report that in vitro preincubation of T cell-depleted BM allografts with a combination of interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) (0.1 microgram/mL each) (n = 8), for 3 days prior to infusion, expands megakaryocyte (MK) precursors. MK-progenitor proliferation was assessed in plasma clot colony assays and liquid cultures following pre-exposure to IL-3/GM-CSF. We observed a 2.8-fold increase in the number of colony-forming units-megakaryocyte (CFU-MK) (17.3 +/- 5.2 vs. 6.1 +/- 3.4) (p = 0.001) and a two-fold increase in burst-forming units-megakaryocyte (BFU-MK) (0.2 vs. 0.1) (p = 0.01) per 2 x 10(5) cells/mL compared to control BM samples cultured for 3 days in medium alone. In secondary cultures, the continued presence of IL-3 and GM-CSF increased the number of CFU-MK by 200-fold (p < 0.0001) over controls and by 9.7-fold over fresh BM. A 33-fold increase (p < 0.0001) in the number of BFU-MK was elicited compared to controls. In addition, IL-3 plus GM-CSF supported increased cellularity within the colonies. The presence of IL-3 or GM-CSF alone resulted in fewer MK colonies and fewer cells per colony than both cytokines combined. In liquid cultures, the percentage of cells expressing platelet glycoprotein (GP) IIb/IIIa in the continued presence of IL-3 and GM-CSF increased following preincubation, yielding a total of 16.0 +/- 2.3 x 10(4) MK/2 x 10(6) cells at day 10 of culture. We propose that ex vivo preincubation with IL-3 and GM-CSF can expand the number of MK precursors and may facilitate platelet recovery post-BMT.
迁延性血小板减少症和出血仍然是骨髓移植（BMT）中的严重并发症。在促进髓系和红系植入方面已取得重大进展，但在加速BMT后血小板生成方面进展甚微。我们报告，在输注前3天，用白细胞介素-3（IL-3）和粒细胞-巨噬细胞集落刺激因子（GM-CSF）（各0.1微克/毫升）（n = 8）的组合对去除T细胞的骨髓同种异体移植物进行体外预孵育，可使巨核细胞（MK）前体细胞扩增。在血浆凝块集落测定和液体培养中，在预先暴露于IL-3/GM-CSF后评估MK祖细胞增殖。我们观察到，与仅在培养基中培养3天的对照骨髓样本相比，每2×10⁵个细胞/毫升中巨核细胞集落形成单位（CFU-MK）的数量增加了2.8倍（17.3±5.2对6.1±3.4）（p = 0.001），巨核细胞爆式集落形成单位（BFU-MK）增加了两倍（0.2对0.1）（p = 0.01）。在传代培养中，IL-3和GM-CSF的持续存在使CFU-MK的数量比对照增加了200倍（p < 0.0001），比新鲜骨髓增加了9.7倍。与对照相比，BFU-MK的数量增加了33倍（p < 0.0001）。此外，IL-3加GM-CSF支持集落内细胞数量增加。单独存在IL-3或GM-CSF时，产生的MK集落比两种细胞因子联合使用时少，且每个集落中的细胞也少。在液体培养中，在持续存在IL-3和GM-CSF的情况下，预先孵育后表达血小板糖蛋白（GP）IIb/IIIa的细胞百分比增加，在培养第10天时每2×10⁶个细胞中产生总共16.0±2.3×10⁴个MK。我们提出，用IL-3和GM-CSF进行体外预孵育可扩大MK前体细胞的数量，并可能促进BMT后血小板的恢复。
