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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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Growth curve of DC2.4 cells. (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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f3-medscimonit-17-5-br125: Growth curve of DC2.4 cells. (A) control group; (B) 50 μg/L Dex group; (C) 100 μg/L Dex group; (D) 200 μg/L Dex group.

Mentions: The growth curve of DC2.4 cells was plotted according to the results of cell counting (Figure 3). The doubling times were 17.04 h, 17.40 h, 16.96 h, and 17.21 h for the control group, 50 μg/L dexamethasone group, 100 μg/L dexamethasone group, and 200 μg/L dexamethasone group, respectively, showing no significant difference in doubling time between dexamethasone-treated groups and the control group. In the first 3 days of culture, the number of cells did not markedly increase and the cells were in the latent phase. Thereafter, the number of cells began to increase markedly, indicating that the cells were in the exponential growth phase. Dexamethasone treatment caused no observable abnormality in the growth of DC2.4 cells. Microscopically, DC2.4 cells in various groups reached nearly full confluence at day 5 and completely covered the culture plates at day 6, when local cell clusters were noted.


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)

Growth curve of DC2.4 cells. (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

f3-medscimonit-17-5-br125: Growth curve of DC2.4 cells. (A) control group; (B) 50 μg/L Dex group; (C) 100 μg/L Dex group; (D) 200 μg/L Dex group.
Mentions: The growth curve of DC2.4 cells was plotted according to the results of cell counting (Figure 3). The doubling times were 17.04 h, 17.40 h, 16.96 h, and 17.21 h for the control group, 50 μg/L dexamethasone group, 100 μg/L dexamethasone group, and 200 μg/L dexamethasone group, respectively, showing no significant difference in doubling time between dexamethasone-treated groups and the control group. In the first 3 days of culture, the number of cells did not markedly increase and the cells were in the latent phase. Thereafter, the number of cells began to increase markedly, indicating that the cells were in the exponential growth phase. Dexamethasone treatment caused no observable abnormality in the growth of DC2.4 cells. Microscopically, DC2.4 cells in various groups reached nearly full confluence at day 5 and completely covered the culture plates at day 6, when local cell clusters were noted.

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