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- Title
Priming effects of tumor necrosis factor-α on production of reactive oxygen species during <i>Toxoplasma gondii</i> stimulation and receptor gene expression in differentiated HL-60 cells.
- Authors
Kikuchi-Ueda, Takane; Ubagai, Tsuneyuki; Ono, Yasuo
- Abstract
Neutrophils are among the principal effector cells that protect against infectious agents, in part by producing reactive oxygen species (ROS) via the actions of tumor necrosis factor-α (TNF-α). In this study, we investigated whether HL-60 cells that had been differentiated into neutrophil-like cells by all- trans retinoic acid could be primed with TNF-α similar to human neutrophils. Our results showed that when differentiated HL-60 (dHL-60) cells were primed with TNF-α for 10 min, ROS production induced by zymosan A or phorbol myristate acetate (PMA) was enhanced in a TNF-α-dose-dependent manner. In addition, when dHL-60 cells were stimulated with live tachyzoites of Toxoplasma gondii after TNF-α priming, ROS production was also enhanced. Thus, dHL-60, similar to neutrophils, produced ROS after PMA, zymosan A, or T. gondii stimulation. Furthermore, we examined gene expression in dHL-60 cells after TNF-α treatment. The pro-inflammatory cytokine IL-6 was up-regulated more than 1.6-fold by 0.1 ng/mL TNF-α. Endogenous TNF-α was down-regulated by priming. IL-8 receptors genes were not affected by priming with 0.1 ng/mL or 1 ng/mL TNF-α. Complement receptor (CR) 1 and CR3 gene expression was not affected by TNF-α priming for 10 min. However, when the priming period was extended to 1 h, CR1 and CR3 genes were up-regulated 1.3 and 1.4-fold, respectively. Expression of the cell-surface CR3 (CD11b) was not significantly affected by TNF-α for 15 min but was slightly enhanced after priming for 2 h. These results suggest that dHL-60 cells may be used as a substitute for neutrophils when evaluating the effects of cytokines or immunomodulator agents.
- Subjects
TUMOR necrosis factors; REACTIVE oxygen species; TOXOPLASMA gondii; GENE expression; CANCER cell differentiation; NEUTROPHILS; REVERSE transcriptase polymerase chain reaction
- Publication
Journal of Infection & Chemotherapy (Springer Science & Business Media B.V.), 2013, Vol 19, Issue 6, p1053
- ISSN
1341-321X
- Publication type
Article
- DOI
10.1007/s10156-013-0619-4