Quesenberry P J, McGrath H E, Williams M E, Robinson B E, Deacon D H, Clark S, Urdal D, McNiece I K
University of Virginia Health Sciences Center, Department of Internal Medicine, School of Medicine, Charlottesville 22908.
Exp Hematol. 1991 Jan;19(1):35-41.
We have previously demonstrated that interleukin 3 (IL-3), granulocyte-macrophage colony-stimulating factor (GM-CSF), and granulocyte colony-stimulating factor (G-CSF) stimulate various aspects of megakaryocytopoiesis. We have investigated the capacity of interleukin 6 (IL-6) to stimulate megakaryocyte colony formation from both normal Balb/C marrow and light-density marrow extensively depleted of adherent, pre-B, B and T cells. Human recombinant IL-6 (167 ng/ml) stimulated megakaryocyte colony formation from normal marrow (8.6 +/- 1 megakaryocyte colony-forming units [CFU-meg]/10(5) cells) as compared to control (1.5 +/- 4 CFU-meg/10(5) cells) in 16 determinations (p less than 0.01). IL-6 (167 ng/ml) also stimulated CFU-meg formation from depleted marrow (control, 10.8 +/- 4 CFU-meg/10(5) cells versus IL-6, 68 +/- 19 CFU-meg/10(5) cells in 12 determinations, p less than 0.01). IL-6 synergistically augmented IL-3-induced colony formation (139% IL-3 control, 120% calculated IL-3 plus IL-6 control, n = 11, p less than 0.01) in normal marrow and showed an additive effect in depleted marrow (133% IL-3 control, p less than 0.01, 114% of IL-3 plus IL-6, value not significant [NS] at 0.05 level). Studies with recombinant murine IL-6 gave similar results. There was an increasing level of megakaryocyte colony-stimulating activity from G-CSF (16,667 U/ml, 2.47 +/- 0.6 CFU-meg/10(5) cells, n = 17), to IL-6 (167 ng/ml, 8.47 +/- 0.96 CFU-meg/10(5) cells, n = 19), to GM-CSF (52 U/ml, 23 +/- 4 CFU-meg/10(5) cells, n = 14), to IL-3 (167 U/ml, 48 +/- 5 CFU-meg/10(5) cells, n = 20) as compared to media-stimulated marrow (range 1.29-1.86 CFU-meg/10(5) cells). A similar hierarchy was seen with depleted marrow. Combinations of factors (including IL-3, GM-CSF, G-CSF, and IL-6) tested against normal unseparated murine marrow did not further augment CFU-meg numbers over IL-3 plus IL-6 but did increase colony size. These data suggest that IL-6 is an important megakaryocyte regulator, that at least four growth factors interact synergistically or additively to regulate megakaryocytopoiesis, and that combinations of growth factors, possibly in physical association, might be critical in stimulating megakaryocyte stem cells.
我们之前已经证明，白细胞介素3（IL-3）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）和粒细胞集落刺激因子（G-CSF）可刺激巨核细胞生成的各个方面。我们研究了白细胞介素6（IL-6）刺激正常Balb/C骨髓以及大量去除贴壁细胞、前B细胞、B细胞和T细胞的低密度骨髓中巨核细胞集落形成的能力。在16次测定中，与对照组（1.5±4个巨核细胞集落形成单位[CFU-meg]/10⁵个细胞）相比，重组人IL-6（167 ng/ml）刺激正常骨髓中的巨核细胞集落形成（8.6±1个CFU-meg/10⁵个细胞）（p<0.01）。在12次测定中，IL-6（167 ng/ml）也刺激去除细胞后的骨髓中的CFU-meg形成（对照组为10.8±4个CFU-meg/10⁵个细胞，IL-6组为68±19个CFU-meg/10⁵个细胞，p<0.01）。在正常骨髓中，IL-6协同增强IL-3诱导的集落形成（IL-3对照组的139%，计算得出的IL-3加IL-6对照组的120%，n = 11，p<0.01），在去除细胞后的骨髓中显示出相加作用（IL-3对照组的133%，p<0.01，IL-3加IL-6的114%，在0.05水平无统计学意义[NS]）。用重组鼠IL-6进行的研究得到了类似结果。从G-CSF（16,667 U/ml，2.47±0.6个CFU-meg/10⁵个细胞，n = 17）到IL-6（167 ng/ml，8.47±0.96个CFU-meg/10⁵个细胞，n = 19），再到GM-CSF（52 U/ml，23±4个CFU-meg/10⁵个细胞，n = 14），最后到IL-3（167 U/ml，48±5个CFU-meg/10⁵个细胞，n = 20），与培养基刺激的骨髓相比（范围为1.29 - 1.86个CFU-meg/10⁵个细胞），巨核细胞集落刺激活性水平逐渐升高。在去除细胞后的骨髓中也观察到了类似的等级关系。针对未分离的正常鼠骨髓测试的多种因子组合（包括IL-3、GM-CSF、G-CSF和IL-6），与IL-3加IL-6相比，并没有进一步增加CFU-meg数量，但增加了集落大小。这些数据表明，IL-6是一种重要的巨核细胞调节因子，至少四种生长因子相互协同或相加作用来调节巨核细胞生成，并且生长因子组合，可能以物理结合的形式，在刺激巨核细胞干细胞方面可能至关重要。
