Tsukada J, Misago M, Kikuchi M, Sato T, Ogawa R, Ota T, Oda S, Morimoto I, Chiba S, Eto S
First Department of Internal Medicine, School of Medicine, University of Occupational and Environmental Health, Fukuoka, Japan.
Blood. 1992 Jul 1;80(1):37-45.
We investigated the interactions between human erythropoietin (hEpo) and serum factor(s) on murine megakaryocyte (MK) colony formation. Serum-free cultures supported the growth of a large number of murine MK colonies in the presence of murine interleukin-3 (mIL-3). The addition of fetal calf serum (FCS) to mIL-3-containing cultures resulted in only a minimal increase in the number of murine MK colonies. In contrast, hEpo alone had no murine MK colony-stimulating activities in serum-free cultures. hEpo required the presence of FCS, murine serum, or human serum in cultures to promote murine MK colony growth and synergized with these sera to stimulate murine MK colony formation. Furthermore, sera from patients with aplastic anemia showed higher synergistic activities with hEpo than sera from hematologically normal persons (normal human serum). When normal human serum was fractionated by gel-filtration chromatography, two peaks with the synergistic activity were observed in the eluent. However, serum did not show any synergistic effects with hEpo on the growth of murine GM colonies or murine colony-forming unit-erythroid-derived colonies. Although human serum synergized with hEpo to stimulate murine MK colony formation, human cytokines such as IL-3, IL-4, IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte-CSF (G-CSF) failed to induce murine MK colony formation in Epo-containing cultures. In cultures containing human IL-1 alpha + human IL-6 + hEpo as well as in cultures containing hEpo, human IL-3 and human GM-CSF failed to show stimulatory effects on murine MK colony formation. Moreover, the synergistic activity of human serum with hEpo could not be neutralized by antibodies such as antihuman IL-1 alpha, antihuman IL-3, antihuman IL-4, antihuman IL-6, antihuman G-CSF, and antihuman GM-CSF. Our data show that serum contains a growth factor(s) that synergizes with Epo to stimulate the proliferation and differentiation of MK precursors, and strongly suggest that this factor(s) is an unique growth factor(s) that is distinct from IL-1 alpha, IL-3, IL-4, IL-6, G-CSF, and GM-CSF.
我们研究了人促红细胞生成素（hEpo）与血清因子对小鼠巨核细胞（MK）集落形成的相互作用。在无血清培养体系中，在小鼠白细胞介素-3（mIL-3）存在的情况下，可支持大量小鼠MK集落的生长。向含mIL-3的培养体系中添加胎牛血清（FCS），小鼠MK集落数量仅略有增加。相比之下，单独的hEpo在无血清培养体系中没有小鼠MK集落刺激活性。hEpo需要在培养体系中存在FCS、小鼠血清或人血清才能促进小鼠MK集落生长，并与这些血清协同刺激小鼠MK集落形成。此外，再生障碍性贫血患者的血清与hEpo的协同活性高于血液学正常者的血清（正常人血清）。当正常人血清通过凝胶过滤色谱法分离时，在洗脱液中观察到两个具有协同活性的峰。然而，血清对小鼠GM集落或小鼠红系集落形成单位衍生的集落的生长与hEpo没有任何协同作用。尽管人血清与hEpo协同刺激小鼠MK集落形成，但人细胞因子如IL-3、IL-4、IL-6、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）在含Epo的培养体系中未能诱导小鼠MK集落形成。在含人IL-1α+人IL-6+hEpo的培养体系以及含hEpo、人IL-3和人GM-CSF的培养体系中，均未显示出对小鼠MK集落形成的刺激作用。此外，人血清与hEpo的协同活性不能被抗人IL-1α、抗人IL-3、抗人IL-4、抗人IL-6、抗人G-CSF和抗人GM-CSF等抗体中和。我们的数据表明，血清中含有一种生长因子，它与Epo协同刺激MK前体细胞的增殖和分化，并强烈提示该因子是一种独特的生长因子，不同于IL-1α、IL-3、IL-4、IL-6、G-CSF和GM-CSF。
