Immune Netw.  2017 Jun;17(3):179-185. 10.4110/in.2017.17.3.179.

Diclofenac Inhibits 27-hydroxycholesterol-induced Differentiation of Monocytic Cells into Mature Dendritic Cells

• Son Y
• Kim BY
• Park YC
• Kim K

• KMID: 2383356
• DOI: http://doi.org/10.4110/in.2017.17.3.179

Abstract

We investigated whether diclofenac could influence the development of antigen-presenting cells in an oxygenated cholesterol-rich environment by determining its effects on the 27-hydroxycholesterol (27OHChol)-induced differentiation of monocytic cells into mature dendritic cells (mDCs). Treatment of human THP-1 monocytic cells with diclofenac antagonized the effects of 27OHChol by attenuating dendrite formation and cell attachment and promoting endocytic function. Diclofenac inhibited the transcription and surface expression of the mDC markers of CD80, CD83, and CD88, and reduced the 27OHChol-induced elevation of surface levels of MHC class I and II molecules to the basal levels in a dose-dependent manner. It also reduced the expression of CD197, a molecule involved in DC homing and migration. These results indicate that diclofenac inhibits the differentiation of monocytic cells into mDCs, thereby potentially modulating adaptive immune responses in a milieu rich in cholesterol oxidation products.

Keyword

27-hydroxycholesterol; Dendritic cell; Diclofenac; Differentiation; Monocytic cell

MeSH Terms

Antigen-Presenting Cells
Cholesterol
Dendrites
Dendritic Cells*
Diclofenac*
Humans
Oxygen
Cholesterol
Diclofenac
Oxygen

Figure

• 

  Figure 1 Effects of diclofenac on cell attachment and morphological changes. (A) After incubation of THP-1 cells (1×106 cells/60-mm culture dish) for 48 h with or without 5, 10, or 25 µg/ml diclofenac in the presence of 27OHChol (2.5 µg/ml), the adherent cells were counted. Data are expressed as mean±SD (n=3 replicates/group). ###p<0.001 vs. control; ***p<0.001 vs. 27OHChol. (B) After incubation with 27OHChol (2.5 µg/ml) for 48 h with or without 5, 10, or 25 µg/ml diclofenac, THP-1 cells were visualized through a microscope. Df, diclofenac.

• 

  Figure 2 Effects of diclofenac on functional changes in monocytic cells induced by 27OHChol. After incubation for 48 h with 27OHChol (2.5 µg/ml) with or without 5, 10, or 25 µg/ml diclofenac, THP-1 cells were treated with 1 mg/ml FITC-conjugated dextran for 1 h. Cells were analyzed by flow cytometry. Results are the representative of three independent experiments. Df, diclofenac.

• 

  Figure 3 Effects of diclofenac on the transcription and surface expression of mDC markers induced by 27OHChol. (A) THP-1 cells (1<106 cells/60-mm culture dish) were cultured for 48 h with or without 5, 10, or 25 µg/ml diclofenac in the presence of 27OHChol. Transcript levels of CD80, CD83, and CD88 genes were analyzed by real-time PCR. Data are expressed as mean±SD (n=3 replicates/group). ###p<0.001 vs. control; ***p<0.001 vs. 27OHChol. (B) THP-1 cells were cultured for 48 h in the presence of 2.5 µg/ml 27OHChol with or without the indicated concentrations of diclofenac. Cells were analyzed by flow cytometry after immunostaining with antibodies against CD80, CD83, and CD88. Results are the representative of three independent experiments. Df, diclofenac.

• 

  Figure 4 Effects of diclofenac on the expression of MHC class I and II molecules induced by 27OHChol. After incubation for 48 h with 27OHChol (2.5 µg/ml) with or without 5, 10, or 25 µg/ml diclofenac, the stimulated THP-1 cells (1×106 cells/60 mm-culture dish) were immunostained for MHC class I and II. Fluorescence was analyzed by flow cytometry. Results are the representative of three independent experiments. Df, diclofenac.

• 

  Figure 5 Diclofenac reduced the expression of the homing molecule CD197 induced by 27OHChol on monocytic cells. THP-1 cells (1×106 cells/60-mm culture dish) were cultured for 48 h with 2.5 mg/ml 27OHChol with or without the indicated concentrations of diclofenac. The harvested cells were immunostained with antibodies against CD197 and analyzed by flow cytometry. Results are the representative of three independent experiments. Df, diclofenac.

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