Premium
Oxygen Metabolism of the HL‐60 Cell Line: Comparison of the Effects of Monocytoid and Neutrophilic Differentiation
Author(s) -
Thompson Blonnie Y.,
Sivam Geetha,
Britigan Bradley E.,
Rosen Gerald M.,
Cohen Myron S.
Publication year - 1988
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.43.2.140
Subject(s) - superoxide , zymosan , cytochalasin b , phagosome , biology , phagocyte , cell culture , biochemistry , microbiology and biotechnology , cytochalasin , myeloperoxidase , phorbol , azurophilic granule , cellular differentiation , metabolism , chemistry , phagocytosis , cell , immunology , inflammation , in vitro , enzyme , genetics , protein kinase c , cytoskeleton , gene
HL‐60 cells are promyelocytic leukemia cells that respond to culture conditions with differentiation into granulocytelike or macrophagelike phagocytes. O 2 metabolism is critical to the microbicidal function of phagocytic cells. O 2 metabolism was studied in HL‐60 cells differentiated with dimethylsulfoxide (Me 2 SO) and 1,25(OH) 2 D 3 , with the objective of 1) determining the validity of these cells as models for human neutrophils and monocytes, respectively, and 2) determining whether these ceils are capable of forming hydroxyl radical. Me 2 SO‐treated cells had morphology consistent with human neutrophils. O 2 consumption by these cells in response to phorbol myristate acetate (PMA; 100 ng/ml) or opsonized zymosan (3 mg/ml) was less than that by neutrophils, as was superoxide formation. O 2 metabolism was not inhibited by KCN or antimycin A. Myeloperoxidase (MPO) activity decreased during differentiation but remained greater than that of human neutrophils. Cytochalasin B enhanced recovery of superoxide secreted in response to zymosan, implying its release from the phagosome. 1,25(OH) 2 D 3 ‐treated cells had morphology consistent with monocytes. O 2 consumption and superoxide release were less than with Me 2 O‐treated cells. Unlike the case with human monocytes, O 2 consumption was not inhibited by KCN or antimycin A. MPO activity was minimally reduced by differentiation. Cytochalasin B inhibited recovery of superoxide. Luminol‐dependent luminescence was greater among 1,25(OH) 2 D 3 ‐treated cells than among Me 2 SO‐treated cells. Free radicals were also measured with a spin trapping technique using 5,5‐dimethyl‐1‐pyrroline‐N‐oxide (DMPO). Spin trapping allows direct, simultaneous detection of superoxide and hydroxyl radicals. Regardless of the mechanism of differentiation, only superoxide was formed by HL‐60 cells. These results show that Me 2 SO‐treated HL‐60 cells represent an excellent model for the study of human neutrophil oxidative function. However, 1,25(OH) 2 D 3 ‐treated cells are quite different in their O 2 metabolism from peripheral blood monocytes.