Migliaccio A R, Migliaccio G, Shimada Y, Adamson J W
Laboratory of Hematopoietic Growth Factors, New York Blood Center, New York 10021.
Biotechnol Ther. 1989;1(4):347-60.
The purpose of this study was to analyze the effects of recombinant human interleukin 4 (IL-4) on the differentiation and proliferation in vitro of human granulocyte/macrophage (GM) and erythroid progenitors. IL-4 was added to either fetal bovine serum (FBS)-supplemented or to FBS-deprived cultures of unfractionated human marrow cells or marrow cells depleted of adherent and/or T cells. Paradoxical effects similar to those reported in the murine system were detected in these experiments. In FBS-supplemented cultures, IL-4, which had no effect on the growth or erythroid bursts (from burst-forming cells; BFU-E) detected in the presence of Epo alone, decreased by 46% the number of erythroid bursts detected in the presence of Epo and phytohemagglutinin-stimulated leukocyte-conditioned medium (PHA-LCM). In contrast, in FBS-deprived cultures, IL-4 increased by 30-700% the number of erythroid bursts in cultures containing Epo alone or containing Epo, IL-3, and GM-CSF. The stimulatory effect of IL-4 on erythroid burst growth under FBS-deprived conditions was particularly evident when adherent cells were removed. Under the conditions investigated, IL-4 had little effect on the growth of GM colonies. In FBS-deprived suspension cultures of nonadherent, T-cell-depleted marrow cells, IL-4 maintained both the number of BFU-E and CFU-GM for at least 8 days. In these cultures, IL-4 antagonized the capacity of IL-3 to increase the number of BFU-E but IL-4 and IL-3 acted together to maintain the number of CFU-GM. To determine if IL-4 acted directly or indirectly, its effects on the growth of factor-dependent subclones of the murine progenitor cell line 32D were analyzed. Three subclones were studied: the original IL-3-dependent clone 32D cl.3, the Epo-dependent erythroid clone 32D Epo-1, and the G-CSF-dependent myeloid clone 32D G-1. IL-4 alone failed to induce colony growth from these cell lines. However, IL-4 inhibited by 25% the number of colonies formed by 32D cl.3 in the presence of IL-3 while increasing by 25% and 25-50% the number of colonies formed by 32D Epo-1 and 32D G-1 in the presence of Epo or G-CSF, respectively. These results indicate that human IL-4, as its murine counterpart, is a multilineage growth factor with paradoxical effects which are mediated by the direct action of IL-4 on progenitor cells.
本研究的目的是分析重组人白细胞介素4（IL-4）对人粒细胞/巨噬细胞（GM）和红系祖细胞体外分化和增殖的影响。将IL-4添加到补充有胎牛血清（FBS）的未分级人骨髓细胞培养物中，或添加到去除贴壁细胞和/或T细胞的FBS缺乏的未分级人骨髓细胞培养物中。在这些实验中检测到了与在小鼠系统中报道的类似的矛盾效应。在补充有FBS的培养物中，IL-4对单独存在促红细胞生成素（Epo）时检测到的生长或红系爆式集落形成单位（BFU-E）无影响，但在Epo和植物血凝素刺激的白细胞条件培养基（PHA-LCM）存在时，可使检测到的红系爆式集落数量减少46%。相反，在FBS缺乏的培养物中，IL-4可使单独含有Epo或含有Epo、IL-3和GM-CSF的培养物中的红系爆式集落数量增加30%-700%。当去除贴壁细胞时，IL-4在FBS缺乏条件下对红系爆式集落生长的刺激作用尤为明显。在所研究的条件下，IL-4对GM集落的生长影响很小。在去除贴壁细胞和T细胞的FBS缺乏的悬浮培养物中，IL-4可使BFU-E和CFU-GM的数量至少维持8天。在这些培养物中，IL-4拮抗IL-3增加BFU-E数量的能力，但IL-4和IL-3共同作用维持CFU-GM的数量。为了确定IL-4是直接还是间接起作用，分析了其对小鼠祖细胞系32D的因子依赖性亚克隆生长的影响。研究了三个亚克隆：原始的IL-3依赖性克隆32D cl.3、Epo依赖性红系克隆32D Epo-1和G-CSF依赖性髓系克隆32D G-1。单独的IL-4未能诱导这些细胞系形成集落。然而，IL-4在IL-存在时可使32D cl.3形成的集落数量减少25%，而在Epo或G-CSF存在时，可使32D Epo-1和32D G-1形成的集落数量分别增加25%和25%-50%。这些结果表明，人IL-4与其小鼠对应物一样，是一种具有矛盾效应的多谱系生长因子，其效应是由IL-4对祖细胞的直接作用介导的。 3
