Kitagawa S, Ohta M, Nojiri H, Kakinuma K, Saito M, Takaku F, Miura Y
J Clin Invest. 1984 Apr;73(4):1062-71. doi: 10.1172/JCI111291.
The alterations of stimulus-induced membrane potential changes, superoxide (O2-)-producing capacity and phagocytic activity during differentiation of human granulocytes were investigated in the human leukemia cell lines HL-60 and KG-1 differentiating in vitro and in human leukemic granulocytes obtained from chronic myelogenous leukemia patients. HL-60 cells incubated with dimethyl sulfoxide or with retinoic acid showed progressively increasing O2- production as well as membrane potential changes (depolarization) on contact with phorbol myristate acetate or the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine, with a concomitant increase in the proportion of mature cells of the granulocytic type. Phagocytosis of latex particles, yeast, and oil droplets appeared 24 h after incubation with dimethyl sulfoxide and anteceded the increment of O2- production and membrane potential changes, both of which appeared concomitantly 3 d after incubation with dimethyl sulfoxide. Similar findings were observed when immature and mature granulocytes obtained from chronic myelogenous leukemia patients were stimulated by phorbol ester, the chemotactic peptide, or calcium ionophore A23187, and the amount of O2- production was parallel to the magnitude of membrane potential changes. HL-60 and KG-1 cells incubated for 1-6 d with phorbol myristate acetate showed neither O2- production nor membrane potential changes on contact with phorbol ester, chemotactic peptide, or A23187, although such cells resembled macrophages morphologically, and their phagocytic activity was significantly increased. O2- production and membrane potential changes in normal granulocytes induced by phorbol ester, chemotactic peptide and A23187 were inhibited by 2-deoxyglucose. These findings indicate that the O2--producing system and the system provoking membrane potential changes may develop concomitantly as human granulocytes mature and differentiate, and that the development of these systems and of phagocytic activity may be independently regulated.
在体外分化的人白血病细胞系HL-60和KG-1以及从慢性粒细胞白血病患者获得的人白血病粒细胞中，研究了人类粒细胞分化过程中刺激诱导的膜电位变化、超氧化物（O2-）产生能力和吞噬活性的改变。用二甲基亚砜或视黄酸孵育的HL-60细胞，在与佛波酯肉豆蔻酸酯或趋化肽N-甲酰甲硫氨酰亮氨酰苯丙氨酸接触时，显示出逐渐增加的O2-产生以及膜电位变化（去极化），同时粒细胞类型的成熟细胞比例增加。用二甲基亚砜孵育24小时后出现对乳胶颗粒、酵母和油滴的吞噬作用，且早于O2-产生和膜电位变化的增加，这两者在与二甲基亚砜孵育3天后同时出现。当用佛波酯、趋化肽或钙离子载体A23187刺激从慢性粒细胞白血病患者获得的未成熟和成熟粒细胞时，观察到类似的结果，并且O2-产生量与膜电位变化幅度平行。用佛波酯肉豆蔻酸酯孵育1 - 6天的HL-60和KG-1细胞，在与佛波酯、趋化肽或A23187接触时既不产生O2-也不发生膜电位变化，尽管这些细胞在形态上类似巨噬细胞，并且它们的吞噬活性显著增加。2-脱氧葡萄糖抑制了佛波酯、趋化肽和A23187诱导的正常粒细胞中的O2-产生和膜电位变化。这些发现表明，随着人类粒细胞成熟和分化，O2-产生系统和引起膜电位变化的系统可能同时发展，并且这些系统和吞噬活性的发展可能受到独立调节。
