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24 April 2011

Experimental study of the mechanism of tolerance induction in dexamethasone-treated dendritic cells

Yu-bo GongABCDEFG, Yi-fei HuangABCDEFG, Yan LiABC, Gen-cheng HanABC, Yu-rong LiABC, Da-jiang WangCDF, Gai-ping DuCDF, Ji-feng YuCDF, Jing SongCDF

DOI: 10.12659/MSM.881758

Med Sci Monit 2011; 17(5): BR125-131

Abstract

Background: The aim of this study was to investigate the mechanisms underlying tolerance induction of dexamethasone (Dex)-treated dendritic cells (DCs).
Material/Methods: Well-grown DC2.4 cells were randomly assigned to receive control, 50 µg/L, 100 µg/L, or 200 µg/L of dexamethasone and then were cultured for 6 days. The expressions of CD80, CD86, galectin-9, and PD-L1 on the surface of DC2.4 cells were analyzed with flow cytometry and the level of IL-12 secreted by DC2.4 cells was determined by ELISA. The stimulating activity of DC2.4 cells on allogeneic T cells was assessed with mixed lymphocyte reaction. Dexamethasone-treated DC2.4 cells were co-cultured with allogeneic splenic lymphocytes and the Foxp3 expression in naive T lymphocytes was determined with flow cytometry.
Results: Compared with the control group, the expressions of CD80, CD86, galectin-9, and PD-L1 on the surface of DC2.4 cells exposed to different doses of dexamethasone showed no significant changes; however, dexamethasone treatment significantly reduced IL-12 secretion and inhibited DC2.4’s stimulation on the proliferation of allogeneic T lymphocytes. Moreover, dexamethasone-treated DC2.4 cells effectively promoted FOXP3 expression in naive T lymphocytes.
Conclusions: DC2.4 is a stable cell line with high expressions of CD80, CD86, and PD-L1. Dexamethasone does not significantly change the cell phenotype of DC2.4 cells, but inhibits the secretion of IL-12 cytokine and attenuates DC2.4’s stimulation of the proliferation of allogeneic T cells. Dexamethasone-treated DC2.4 cells also effectively promote FOXP3 expression in naive T lymphocytes.

Keywords: Interleukin-12 - metabolism, Immune Tolerance - drug effects, Galectins - metabolism, Forkhead Transcription Factors - metabolism, Dexamethasone - pharmacology, Dendritic Cells - immunology, Cell Shape - drug effects, Cell Proliferation - drug effects, Cell Line, Cell Adhesion - drug effects, Antigens, CD86 - metabolism, Antigens, CD80 - metabolism, Lymphocyte Culture Test, Mixed, Subcellular Fractions - metabolism, Time Factors

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750