Straneva J E, Briddell R A, McDonald T P, Yang H H, Hoffman R
Department of Medicine, Indiana University School of Medicine, Indianapolis.
Exp Hematol. 1989 Dec;17(11):1122-7.
Aplastic anemia serum (AAS) contains humoral factors that alter both proliferation and maturation of human megakaryocytes (MK). The ability of AAS to augment MK colony formation (colony-forming unit, CFU-MK) was neutralized by an antiserum against MK colony-stimulating factor (MK-CSF), a glycoprotein isolated from AAS. The adsorbed AAS still retained the ability to accelerate cytoplasmic maturation of recognizable MK. Similar experiments were done with thrombocytopoiesis-stimulating factor (TSF) and an anti-TSF antiserum to further define the activity in AAS responsible for accelerating cytoplasmic maturation. Bone marrow fractions enriched for recognizable human MK, but devoid of CFU-MK, were obtained by centrifugal elutriation and placed in short-term liquid cultures. MK progressed through identifiable maturation stages (1-4) more quickly in the presence of either TSF or AAS. TSF slightly enhanced the cloning efficiencies of CFU-MK, but did not alter the number of MK in individual colonies derived from non-adherent, low-density, T-cell-depleted bone marrow. In contrast, granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin 3 (IL-3), and crude AAS substantially augmented both MK colony formation and cells per colony. TSF also doubled the percent 35S incorporation into platelets of immunothrombocythemic mice, but stimulation was completely abolished by anti-TSF. Anti-TSF antiserum was then used to analyze the promotion of MK colony formation by cytokines. Cloning efficiencies of CFU-MK were reduced to baseline values when TSF was pretreated with anti-TSF; however, the MK colony-stimulating activity (MK-CSA) of GM-CSF, IL-3, or AAS was not altered by adsorption with anti-TSF. In contrast, the cytoplasmic maturation of recognizable MK was slower, and fewer mature stage-4 cells were present at days 1-3 in AAS adsorbed with anti-TSF than MK cultured in AAS treated with normal rabbit serum or untreated AAS. Therefore, TSF appears to be a major factor in AAS that accelerates terminal maturation of human MK. TSF primarily affects megakaryocytopoiesis by promoting MK maturation rather than enhancing CFU-MK proliferation.
再生障碍性贫血血清（AAS）含有可改变人巨核细胞（MK）增殖和成熟的体液因子。AAS增强MK集落形成（集落形成单位，CFU-MK）的能力被一种针对MK集落刺激因子（MK-CSF）的抗血清中和，MK-CSF是一种从AAS中分离出的糖蛋白。吸附后的AAS仍保留加速可识别MK细胞质成熟的能力。用血小板生成刺激因子（TSF）和抗TSF抗血清进行了类似实验，以进一步确定AAS中负责加速细胞质成熟的活性。通过离心淘析获得富含可识别的人MK但不含CFU-MK的骨髓部分，并将其置于短期液体培养中。在TSF或AAS存在的情况下，MK更快地经历可识别的成熟阶段（1-4）。TSF略微提高了CFU-MK的克隆效率，但未改变来自非贴壁、低密度、T细胞耗竭骨髓的单个集落中的MK数量。相比之下，粒细胞-巨噬细胞集落刺激因子（GM-CSF）、白细胞介素3（IL-3）和粗制AAS显著增强了MK集落形成和每个集落中的细胞数量。TSF还使免疫性血小板减少小鼠血小板中35S掺入百分比增加了一倍，但抗TSF完全消除了这种刺激。然后用抗TSF抗血清分析细胞因子对MK集落形成的促进作用。当TSF用抗TSF预处理时，CFU-MK的克隆效率降至基线值；然而，GM-CSF、IL-3或AAS的MK集落刺激活性（MK-CSA）不会因用抗TSF吸附而改变。相比之下，可识别MK的细胞质成熟较慢，与用正常兔血清处理的AAS或未处理的AAS培养的MK相比，用抗TSF吸附的AAS在第1-3天出现的成熟4期细胞更少。因此，TSF似乎是AAS中加速人MK终末成熟的主要因子。TSF主要通过促进MK成熟而非增强CFU-MK增殖来影响巨核细胞生成。
