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

Gong YB, Huang YF, Li Y, Han GC, Li YR, Wang DJ, Du GP, Yu JF, Song J - Med. Sci. Monit. (2011)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Chinese PLA General Hospital, Beijing, China.

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.

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Flow cytometry showing the expression of CD86. (A) control group; (B) 50 μg/L Dex group; (C) 100 μg/L Dex group; (D) 200 μg/L Dex group.
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f5-medscimonit-17-5-br125: Flow cytometry showing the expression of CD86. (A) control group; (B) 50 μg/L Dex group; (C) 100 μg/L Dex group; (D) 200 μg/L Dex group.

Mentions: The percentage of DC2.4 cells that expressed CD86 was 96.46±2.31% in the control group, 95.98±1.96% in the 50 μg/L dexamethasone group, 98.28±1.44% in the 100 μg/L dexamethasone group, and 96.88±1.45% in the 200 μg/L dexamethasone group (Figure 5).


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

Gong YB, Huang YF, Li Y, Han GC, Li YR, Wang DJ, Du GP, Yu JF, Song J - Med. Sci. Monit. (2011)

Flow cytometry showing the expression of CD86. (A) control group; (B) 50 μg/L Dex group; (C) 100 μg/L Dex group; (D) 200 μg/L Dex group.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3539585&req=5

f5-medscimonit-17-5-br125: Flow cytometry showing the expression of CD86. (A) control group; (B) 50 μg/L Dex group; (C) 100 μg/L Dex group; (D) 200 μg/L Dex group.
Mentions: The percentage of DC2.4 cells that expressed CD86 was 96.46±2.31% in the control group, 95.98±1.96% in the 50 μg/L dexamethasone group, 98.28±1.44% in the 100 μg/L dexamethasone group, and 96.88±1.45% in the 200 μg/L dexamethasone group (Figure 5).

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Chinese PLA General Hospital, Beijing, China.

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.

Show MeSH
Related in: MedlinePlus