Functional maturation of membrane potential changes and superoxide-producing capacity during differentiation of human granulocytes.

Abstract

Abstract. The alterations of stimulus-induced membrane potential changes, superoxide (O°)-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 oflatex particles, yeast, and oil droplets appeared 24 h after incubation with dimethyl sulfoxide and anteceded the increment of02 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 A23 187, and the amount of 0°production was parallel to the magnitude of membrane potential changes. HL-60 and KG-1 cells incubated for 1-6 d with phorbol Address correspondence to Dr. Kitagawa, Department of Pediatrics, National Jewish Hospital and Research Center, University of Colorado School of Medicine, Denver, CO 80206 (until August 1985).Received for publication 19 April 1983 and in revised form 22 December 1983. myristate acetate showed neither 0°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. 02-production and membrane potential changes in normal granulocytes induced by phorbol ester, chemotactic peptide and A23 187 were inhibited by 2-deoxyglucose. These findings indicate that the 0--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.

Stimulation by leukotriene D4 of increases in the cytosolic concentration of calcium in dimethylsulfoxide-differentiated HL-60 cells.

“…The induction by DMSO of differentiation of HL-60 cells into mature granulocytes with receptors for fMLP and LTB4 has been well-documented (6,11,(19)(20)(21). LTC4 and LTD4 have been shown to enhance human PMN leukocyte adherence to synthetic surfaces in vitro (8) and to microvascular endothelium in vivo (9).…”

Ganglioside GM3: an acidic membrane component that increases during macrophage-like cell differentiation can induce monocytic differentiation of human myeloid and monocytoid leukemic cell lines HL-60 and U937.

“…Functional differentiation was assessed on the basis of phagocytic activity of the cells. Phagocytic activity of undifferentiated and differentiation-induced cells was measured by counting the number of cells that phagocytosed latex or yeast particles as described (25). Polystyrene latex particles (Dow Chemical, Indianapolis; 1.09-,um diameter) were added to the cell suspension in RPMI 1640 medium and were left in contact with the cells for 60 min at 37°C.…”

Expression of the inhibitor of apoptosis (IAP) family members in human neutrophils: up-regulation of cIAP2 by granulocyte colony-stimulating factor and overexpression of cIAP2 in chronic neutrophilic leukemia

“…Human neutrophils and mononuclear cells were prepared from healthy adult donors as described previously, 21 using dextran sedimentation, centrifugation with Conray-Ficoll, and hypotonic lysis of contaminated erythrocytes. Mature neutrophils were also prepared from a patient with CNL, 6 patients with chronic myelogenous leukemia (CML), and 4 healthy donors receiving G-CSF administration for allogeneic peripheral blood stem cell transplantation using the same procedure described in the previous sentence.…”

“…Mature neutrophils were also prepared from a patient with CNL, 6 patients with chronic myelogenous leukemia (CML), and 4 healthy donors receiving G-CSF administration for allogeneic peripheral blood stem cell transplantation using the same procedure described in the previous sentence. 21 Informed consent was obtained from all subjects. Monocytes and lymphocytes were purified from mononuclear cells by centrifugal elutriation in a Hitachi SRR6Y elutriation rotor (Hitachi, Tokyo, Japan), as described previously.…”

“…21,22 The final concentration of cytochrome c was 100 M, and the final cell concentration was 1 ϫ 10 6 cells/mL. The amount of O 2 Ϫ release was calculated from cytochrome c reduced for 5 minutes after the addition of FMLP (10 Ϫ7 M).…”