Departments of
* Biochemistry and
Medicine, Division of Hematology, and
Division of Pathology, Juntendo University, School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan
Correspondence: Isao Nagaoka, Department of Biochemistry, Juntendo University, School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan. E-mail: nagaokai{at}med.juntendo.ac.jp
To understand the expression of NADPH oxidase components during neutrophil maturation, we examined the expression of mRNAs and proteins for NADPH oxidase components, and the superoxide-producing activity using HL-60 cells incubated with dimethyl sulfoxide (DMSO). Northern blot and Western blot analyses revealed that gp91phox, p67phox, and p47phox were expressed after myelocyte stages, whereas p22phox, p40phox, and rac-2 were expressed from the promyelocyte stage. Furthermore, immunocytochemical staining of DMSO-induced HL-60 cells indicated that gp91phox, p67phox, and p47phox were detected only after myelocyte stages (myelocytes, metamyelocytes, band cells, and segmented cells), whereas p22phox, p40phox, and rac-2 were detected from the promyelocyte stage. In addition, nitro blue tetrazolium (NBT) assay showed that superoxide could be produced after myelocyte stages but not produced before promyelocyte stages. Moreover, almost the same results as those with DMSO-induced HL-60 cells were obtained using human bone-marrow cells by immunocytochemical staining and NBT assay, except that p22phox was detected by immunocytochemical staining after myelocyte stages in bone-marrow cells. Together, these observations indicate that all the components for NADPH oxidase are expressed, and the superoxide-producing activity is obtained after myelocyte stages during neutrophil maturation.
Key Words: superoxide • cytochrome b558 • cytosolic factor
This article has been cited by other articles:
 |
|
 |
|
  M. C. B. Ammons, D. W. Siemsen, L. K. Nelson-Overton, M. T. Quinn, and K. A. Gauss Binding of Pleomorphic Adenoma Gene-like 2 to the Tumor Necrosis Factor (TNF)-{alpha}-responsive Region of the NCF2 Promoter Regulates p67phox Expression and NADPH Oxidase Activity J. Biol. Chem., June 15, 2007; 282(24): 17941 - 17952. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. J. Spellberg, M. Collins, V. Avanesian, M. Gomez, J. E. Edwards Jr., C. Cogle, D. Applebaum, Y. Fu, and A. S. Ibrahim Optimization of a myeloid cell transfusion strategy for infected neutropenic hosts J. Leukoc. Biol., March 1, 2007; 81(3): 632 - 641. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. J. Burkitt and A. Raafat Nitric oxide generation from hydroxyurea: significance and implications for leukemogenesis in the management of myeloproliferative disorders Blood, March 15, 2006; 107(6): 2219 - 2222. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. J. O'Brien, C. Krema, T. Heimann, and H. Zhao Expression of NADPH oxidase in rabbit corneal epithelial and stromal cells in culture. Invest. Ophthalmol. Vis. Sci., March 1, 2006; 47(3): 853 - 863. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. A. Bezdecny, R. A. Roth, and P. E. Ganey Effects of 2,2',4,4'-Tetrachlorobiphenyl on Granulocytic HL-60 Cell Function and Expression of Cyclooxygenase-2 Toxicol. Sci., April 1, 2005; 84(2): 328 - 334. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. A. Gauss, P. L. Bunger, T. C. Larson, C. J. Young, L. K. Nelson-Overton, D. W. Siemsen, and M. T. Quinn Identification of a novel tumor necrosis factor {alpha}-responsive region in the NCF2 promoter J. Leukoc. Biol., February 1, 2005; 77(2): 267 - 278. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. T. Quinn and K. A. Gauss Structure and regulation of the neutrophil respiratory burst oxidase: comparison with nonphagocyte oxidases J. Leukoc. Biol., October 1, 2004; 76(4): 760 - 781. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. G. Lieber, S. Webb, B. T. Suratt, S. K. Young, G. L. Johnson, G. M. Keller, and G. S. Worthen The in vitro production and characterization of neutrophils from embryonic stem cells Blood, February 1, 2004; 103(3): 852 - 859. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. Teufelhofer, R.-M. Weiss, W. Parzefall, R. Schulte-Hermann, M. Micksche, W. Berger, and L. Elbling Promyelocytic HL60 Cells Express NADPH Oxidase and Are Excellent Targets in a Rapid Spectrophotometric Microplate Assay for Extracellular Superoxide Toxicol. Sci., December 1, 2003; 76(2): 376 - 383. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C.-H. Woo, M.-H. Yoo, H.-J. You, S.-H. Cho, Y.-C. Mun, C.-M. Seong, and J.-H. Kim Transepithelial Migration of Neutrophils in Response to Leukotriene B4 Is Mediated by a Reactive Oxygen Species-Extracellular Signal-Regulated Kinase-Linked Cascade J. Immunol., June 15, 2003; 170(12): 6273 - 6279. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Arroyo, M. Modriansky, F. B. Serinkan, R. I. Bello, T. Matsura, J. Jiang, V. A. Tyurin, Y. Y. Tyurina, B. Fadeel, and V. E. Kagan NADPH Oxidase-dependent Oxidation and Externalization of Phosphatidylserine during Apoptosis in Me2SO-differentiated HL-60 Cells. ROLE IN PHAGOCYTIC CLEARANCE J. Biol. Chem., December 13, 2002; 277(51): 49965 - 49975. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M.-J. Rabiet, M. Tardif, L. Braun, and F. Boulay Inhibitory effects of a dominant-interfering form of the Rho-GTPase Cdc42 in the chemoattractant-elicited signaling pathways leading to NADPH oxidase activation in differentiated HL-60 cells Blood, August 13, 2002; 100(5): 1835 - 1844. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Iwabuchi and I. Nagaoka Lactosylceramide-enriched glycosphingolipid signaling domain mediates superoxide generation from human neutrophils Blood, July 30, 2002; 100(4): 1454 - 1464. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-L. Li, A. J. Valente, M. Qiang, W. Schlegel, M. Gamez, and R. A. Clark Multiple PU.1 sites cooperate in the regulation of p40phox transcription during granulocytic differentiation of myeloid cells Blood, May 29, 2002; 99(12): 4578 - 4587. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-M. Li and A. M. Shah Intracellular Localization and Preassembly of the NADPH Oxidase Complex in Cultured Endothelial Cells J. Biol. Chem., May 24, 2002; 277(22): 19952 - 19960. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Chi, S. Tridandapani, W. Zhong, K. M. Coggeshall, and R. F. Mortensen C-Reactive Protein Induces Signaling Through Fc{gamma}RIIa on HL-60 Granulocytes J. Immunol., February 1, 2002; 168(3): 1413 - 1418. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. A. Gauss, P. L. Bunger, and M. T. Quinn AP-1 is essential for p67phox promoter activity J. Leukoc. Biol., January 1, 2002; 71(1): 163 - 172. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-L. Li, A. J. Valente, L. Wang, M. J. Gamez, and R. A. Clark Transcriptional Regulation of the p67phox Gene. ROLE OF AP-1 IN CONCERT WITH MYELOID-SPECIFIC TRANSCRIPTION FACTORS J. Biol. Chem., October 12, 2001; 276(42): 39368 - 39378. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Hasebe, J. Hua, A. Someya, P. Morain, F. Checler, and I. Nagaoka Involvement of cytosolic prolyl endopeptidase in degradation of p40-phox splice variant protein in myeloid cells J. Leukoc. Biol., June 1, 2001; 69(6): 963 - 968. [Abstract] [Full Text] [PDF]
|
|
