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. 2011 May 18;13(3):R77.
doi: 10.1186/ar3339.

Histone deacetylase inhibition alters dendritic cells to assume a tolerogenic phenotype and ameliorates arthritis in SKG mice

Kenta Misaki  1 , Akio Morinobu, Jun Saegusa, Shimpei Kasagi, Masaaki Fujita, Yoshiaki Miyamoto, Fumichika Matsuki, Shunichi Kumagai
Affiliations

Histone deacetylase inhibition alters dendritic cells to assume a tolerogenic phenotype and ameliorates arthritis in SKG mice

Kenta Misaki et al. Arthritis Res Ther. .

Abstract

Introduction: The purpose of this study was to elucidate the effects of histone deacetylase inhibition on the phenotype and function of dendritic cells and on arthritis in SKG mice.

Methods: Arthritis was induced in SKG mice by zymosan A injection. Trichostatin A, a histone deacetylase inhibitor, was administered and its effects on arthritis were evaluated by joint swelling and histological evaluation. Interleukin-17 production in lymph node cells was determined by an enzyme-linked immunosorbent assay (ELISA). Foxp3 expression in lymph node cells and the phenotypes of splenic dendritic cells were examined by fluorescence-activated cell sorting (FACS). Bone marrow-derived dendritic cells (BM-DC) were generated with granulocyte macrophage colony-stimulating factor. The effects of trichostatin A on cell surface molecules, cytokine production, indoleamine 2,3-dioxygenase (IDO) expression and T cell stimulatory capacity were examined by FACS, ELISA, quantitative real-time polymerase chain reaction and Western blot, and the allo-mixed lymphocyte reaction, respectively.

Results: Trichostatin A, when administered before the onset of arthritis, prevented SKG mice from getting arthritis. Trichostatin A treatment also showed therapeutic effects on arthritis in SKG mice, when it was administered after the onset of arthritis. Trichostatin A treatment reduced Th17 cells and induced regulatory T cells in lymph node, and also decreased co-stimulatory molecule expression on splenic dendritic cells in vivo. In vitro, trichostatin A markedly suppressed zymosan A-induced interleukin-12 and interleukin-6 production by BM-DC and up-regulated IDO expression at mRNA and protein levels. Trichostatin A-treated BM-DC also showed less T cell stimulatory capacity.

Conclusions: Histone deacetylase inhibition changes dendritic cells to a tolerogenic phenotype and ameliorates arthritis in SKG mice.

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Figures

Figure 1
Figure 1
The effects of trichostatin A on SKG mice. Zymosan A was administered to SKG mice. Dimethyl sulfoxide or trichostatin A was injected subcutaneously daily for 28 days (Day 14 through Day 42). The clinical arthritis scores were evaluated and the results are expressed as the mean ± SE (DMSO group: n = 5, TSA group: n = 5). * P < 0.05.
Figure 2
Figure 2
Histological analysis of SKG mice on Day 35 (after 21 days of treatment). (A) The histological analysis was performed on their hind paw sections stained by hematoxylin and eosin. Tissues are shown at ×40 magnification. Representative results are shown. (B) Histological arthritis scores between the dimethyl sulfoxide- and trichostatin A-treated groups of SKG mice. Mice were killed on Day 35 (treatment for 21 days) and the histological arthritis scores were calculated on their left hind paw. Results are expressed as the mean ± SE (DMSO group: n = 8, TSA group: n = 8, P = 0.0004). CAS, clinical arthritis scores.
Figure 3
Figure 3
Production of IL-17A and expression of Foxp3 by inguinal lymph node cells of SKG mice. (A) Inguinal lymph node cells of SKG mice in each group were collected on Day 35. Cells were stimulated with phorbol myristate acetate/ionomycin and the supernatants were collected after 8 h for the measurement of IL-17A. Values are presented as the mean ± SE (DMSO group: n = 3, TSA group: n = 3, P < 0.0001). (B) The expression of Foxp3 in inguinal lymph node cells in SKG mice. Inguinal lymph node cells of SKG mice were collected on Day 35 in each group as previously described. Cells were stained for anti-CD4, anti-CD25, and Foxp3. The percentage of CD4+CD25+Foxp3+ cells among gated CD4+ cells was determined. Results are expressed as the mean ± SE (DMSO group: n = 4, TSA group: n = 4, P = 0.038).
Figure 4
Figure 4
MHC class II, CD86, CD80, and CD40 expression on CD8α+ splenic conventional dendritic cells. Spleen cells were isolated from dimethyl sulfoxide- or triclostatin A-treated SKG mice on Day 35 and stained for each marker. Gates were set on CD8α+ conventional dendritic cells and cell surface molecules were analyzed on fluorescence-activated cell sorting. (A) Representative results of four experiments are shown by mean fluorescence intensity. (B) The mean fluorescence intensities of indicated molecules in each group were compared. Results are expressed as the mean ± SE (DMSO group: n = 4, TSA group: n = 4, P-value was N.S. in MHC class II, P = 0.035 in CD80, P = 0.023 in CD86, P = 0.012 in CD40). N.S., not significant.
Figure 5
Figure 5
The effects of trichostatin A on bone marrow-derived dendritic cells. (A) The effects of trichostatin A on cytokine production by bone marrow-derived dendritic cells. Bone marrow-derived dendritic cells were generated from SKG mice and stimulated for 18 h with zymosan A and/or triclostatin A. The concentrations of IL-12p70, IL-12p40, and IL-6 in the supernatant were measured by an enzyme-linked immunosorbent assay. Values are presented as the mean ± SE (n = 3). Data are representative of two (IL-12p70) or three (IL-12p40 and IL-6) independent experiments with similar results (P = 0.027 in IL-12p70, P = 0.024 in IL-12p40, P = 0.029 in IL-6). (B) The effects of triclostatin A on indoleamine 2,3-dioxygenase mRNA expression by bone marrow-derived dendritic cells. Bone marrow-derived dendritic cells were generated from SKG mice and stimulated for 18 h with zymosan A (5 μg/ml) and/or trichostatin A (20 nM). IDO1 and IDO2 mRNA expression was measured by quantitative real-time polymerase chain reaction. Representative results of two independent experiments are shown. (C) The effect of trichostatin A on indoleamine 2,3-dioxygenase production by bone marrow-derived dendritic cells. Bone marrow-derived dendritic cells were stimulated with zymosan A (5 μg/ml) and/or trichostatin A (20 nM) for 48 h. Cell lysates were analyzed by Western blotting with anti- indoleamine 2,3-dioxygenase antibodies. The blot is representative of two independent experiments.
Figure 6
Figure 6
The effects of trichostatin A on the phenotype of bone marrow-derived dendritic cells. Bone marrow-derived dendritic cells were generated from SKG mice and incubated for 48 h with zymosan A and/or trichostatin A. Bone marrow-derived dendritic cells were stained for anti-MHC class II, anti-CD54, anti-CD86, anti-CD80, and anti-CD40. (A) The fluorescence activated cell sorting was shown by mean fluorescence intensity. Data are representative of three independent experiments. (B) The mean fluorescence intensity of each group was compared. Results are expressed as the mean ± SE of three independent experiments (P-values were N.S. in MHC class II, CD54 and CD80, P = 0.012 in CD86, P = 0.034 in CD40). N.S, not significant.
Figure 7
Figure 7
The effects of trichostatin A on the T cell stimulatory capacity of dendritic cells. Bone marrow-derived dendritic cells from SKG mice were treated with zymosan A and/or trichostatin A for 18 h, extensively washed, and used for the allo-mixed lymphocyte reaction to assess the T cell stimulatory capacity. Results are expressed as the mean ± SE of four independent experiments (P < 0.01).
Figure 8
Figure 8
Therapeutic effects of trichostatin A after the onset of arthritis in SKG mice. SKG mice were treated with dimethyl sulfoxide or trichostatin A on Day 22, when the mean clinical arthritis score was nearly 1.0 for 24 days. Results are expressed as the mean ± SE (DMSO group: n = 8, TSA group: n = 8). * P < 0.05. N.S., not significant.
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References

    1. Firestein GS. Evolving concepts of rheumatoid arthritis. Nature. 2003;423:356–361. doi: 10.1038/nature01661. - DOI - PubMed
    1. Lutzky V, Hannawi S, Thomas R. Cells of the synovium in rheumatoid arthritis. Dendritic cells. Arthritis Res Ther. 2007;9:219. doi: 10.1186/ar2200. - DOI - PMC - PubMed
    1. Miossec P. Dynamic interactions between T cells and dendritic cells and their derived cytokines/chemokines in the rheumatoid synovium. Arthritis Res Ther. 2008;10:S2. - PMC - PubMed
    1. Santiago-Schwarz F, Anand P, Liu S, Carsons SE. Dendritic cells (DCs) in rheumatoid arthritis (RA): progenitor cells and soluble factors contained in RA synovial fluid yield a subset of myeloid DCs that preferentially activate Th1 inflammatory-type responses. J Immunol. 2001;167:1758–1768. - PubMed
    1. Miossec P. Interleukin-17 in rheumatoid arthritis: if T cells were to contribute to inflammation and destruction through synergy. Arthritis Rheum. 2003;48:594–601. doi: 10.1002/art.10816. - DOI - PubMed

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