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Ubiquitous Over-Expression of Chromatin Remodeling Factor SRG3 Ameliorates the T Cell-Mediated Exacerbation of EAE by Modulating the Phenotypes of both Dendritic Cells and Macrophages.

Lee SW, Park HJ, Jeon SH, Lee C, Seong RH, Park SH, Hong S - PLoS ONE (2015)

Bottom Line: We found that the outcome of EAE development was completely different depending on the expression pattern of SRG3.SRG3 over-expression not only reduced pro-inflammatory cytokine production by DCs but also shifted macrophages from the inducible nitric oxide synthase (iNOS)-expressing M1 phenotype to the arginase-1-expressing M2 phenotype during EAE.Taken together, our results provide the first evidence supporting the development of a new therapeutic strategy for EAE involving the modulation of SRG3 expression to induce M2 and Th2 polarization, thereby inhibiting inflammatory immune responses.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Bioscience and Biotechnology, Institute of Bioscience, Sejong University, Seoul 143-747, Korea; School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Korea.

ABSTRACT
Although SWI3-related gene (SRG3), a chromatin remodeling factor, is critical for various biological processes including early embryogenesis and thymocyte development, it is unclear whether SRG3 is involved in the differentiation of CD4+ T cells, the key mediator of adaptive immune responses. Because it is known that experimental autoimmune encephalomyelitis (EAE) development is determined by the activation of CD4+ T helper cells, here, we investigated the role of SRG3 in EAE development using SRG3 transgenic mouse models exhibiting two distinct SRG3 expression patterns: SRG3 expression driven by either the CD2 or β-actin promoter. We found that the outcome of EAE development was completely different depending on the expression pattern of SRG3. The specific over-expression of SRG3 using the CD2 promoter facilitated EAE via the induction of Th1 and Th17 cells, whereas the ubiquitous over-expression of SRG3 using the β-actin promoter inhibited EAE by promoting Th2 differentiation and suppressing Th1 and Th17 differentiation. In addition, the ubiquitous over-expression of SRG3 polarized CD4+ T cell differentiation towards the Th2 phenotype by converting dendritic cells (DCs) or macrophages to Th2 types. SRG3 over-expression not only reduced pro-inflammatory cytokine production by DCs but also shifted macrophages from the inducible nitric oxide synthase (iNOS)-expressing M1 phenotype to the arginase-1-expressing M2 phenotype during EAE. In addition, Th2 differentiation in β-actin-SRG3 Tg mice during EAE was associated with an increase in the basophil and mast cell populations and in IL4 production. Furthermore, the increased frequency of Treg cells in the spinal cord of β-actin-SRG3 Tg mice might induce the suppression of and accelerate the recovery from EAE symptoms. Taken together, our results provide the first evidence supporting the development of a new therapeutic strategy for EAE involving the modulation of SRG3 expression to induce M2 and Th2 polarization, thereby inhibiting inflammatory immune responses.

No MeSH data available.


Related in: MedlinePlus

SRG3 over-expression driven by the β-actin promoter reduced cytokine production in DCs following LPS stimulation.Flt3L-cultured BMDCs (A) from both WT and CD2-SRG3 Tg B6 mice and (B) from both WT and β-actin-SRG3 Tg B6 mice were stimulated with either vehicle or LPS (40, 200, or 1000 ng/ml) for 14 hrs, and subsequently, the percentage of the IL12p40-expressing population among the CD11c+ BMDCs was assessed via flow cytometry. Representative data from three independent experiments are shown (upper panel). The graph in lower panel represents mean percentage ± SD for the proportion of IL12p40 (n = 3; **P<0.01). (C) WT, β-actin-SRG3 Tg, and CD2-SRG3 Tg mice were i.p. injected with LPS (2 μg) or vehicle. Sixteen hrs later, intracellular TNFα, IL12p40, iNOS, and IL10 production was assessed in splenic CD11c+ DCs via flow cytometry. The means ± SD are shown in the graphs (n = 4; *P<0.05, **P<0.01).
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pone.0132329.g001: SRG3 over-expression driven by the β-actin promoter reduced cytokine production in DCs following LPS stimulation.Flt3L-cultured BMDCs (A) from both WT and CD2-SRG3 Tg B6 mice and (B) from both WT and β-actin-SRG3 Tg B6 mice were stimulated with either vehicle or LPS (40, 200, or 1000 ng/ml) for 14 hrs, and subsequently, the percentage of the IL12p40-expressing population among the CD11c+ BMDCs was assessed via flow cytometry. Representative data from three independent experiments are shown (upper panel). The graph in lower panel represents mean percentage ± SD for the proportion of IL12p40 (n = 3; **P<0.01). (C) WT, β-actin-SRG3 Tg, and CD2-SRG3 Tg mice were i.p. injected with LPS (2 μg) or vehicle. Sixteen hrs later, intracellular TNFα, IL12p40, iNOS, and IL10 production was assessed in splenic CD11c+ DCs via flow cytometry. The means ± SD are shown in the graphs (n = 4; *P<0.05, **P<0.01).

Mentions: DCs play an important role in T cell polarization in the periphery and in the CNS. To determine whether the over-expression of SRG3 has any influence on DC activation elicited by LPS stimulation, BMDCs were generated from CD2-SRG3 Tg, β-actin-SRG3 Tg, and wild type (WT) B6 mice in the presence of Flt3L. After BMDCs were stimulated with LPS, their activation status was monitored based on their IL12p40 cytokine expression pattern. BMDCs from CD2-SRG3 Tg mice produced comparable levels of IL12p40 than those from WT B6 mice (Fig 1A). However, Intriguingly, BMDCs from β-actin-SRG3 Tg mice produced much lower levels of IL12p40 than those from WT B6 mice (Fig 1B), which suggested that the over-expression of SRG3 in DCs restrained the activation of DCs by reducing the production of pro-inflammatory cytokines such as IL12p40. Next, to determine whether SRG3 over-expression affects DC activation by LPS in vivo, β-actin-SRG3 Tg, CD2-SRG3 Tg, and WT B6 mice were i.p. injected with LPS (2 μg) or vehicle. Sixteen hrs later, the expression of TNFα, IL12p40, inducible nitric oxide synthase (iNOS), and IL10 in splenic DCs was examined via flow cytometric analysis. Strikingly, the expression levels of TNFα, IL12p40, and iNOS were decreased by SRG3 over-expression driven by the β-actin promoter. However, as expected, SRG3 over-expression driven by the CD2 promoter did not affect cytokine production in DCs because DCs from CD2-SRG3 Tg mice do not over-express SRG3, as these cells lack CD2 expression. In the vehicle-injected groups, DCs from β-actin-SRG3, but not CD2-SRG3 Tg mice expressed a much lower level of IL12p40 and iNOS but comparable levels of TNFα and IL10 compared to those from WT B6 mice, indicating that SRG3 over-expression in DCs maintains a low basal level of pro-inflammatory gene expression. In addition, no change in IL10 expression was observed between WT B6 mice and either CD2-SRG3 or β-actin-SRG3 Tg mice (Fig 1C; S1 Fig). As expected, the expression levels of SRG3 in splenic CD4+ T cells from both CD2-SRG3 Tg and β-actin-SRG3 Tg mice were up-regulated compared to those from WT B6 mice, whereas the expression levels of SRG3 in splenic DCs were up-regulated in β-actin-SRG3 Tg but not in CD2-SRG3 Tg mice compared to those from WT B6 mice (Fig 1D). These data demonstrated that SRG3 over-expression driven by the β-actin promoter, but not the CD2 promoter, led to the decreased expression of pro-inflammatory cytokines upon LPS stimulation both in vivo and in vitro.


Ubiquitous Over-Expression of Chromatin Remodeling Factor SRG3 Ameliorates the T Cell-Mediated Exacerbation of EAE by Modulating the Phenotypes of both Dendritic Cells and Macrophages.

Lee SW, Park HJ, Jeon SH, Lee C, Seong RH, Park SH, Hong S - PLoS ONE (2015)

SRG3 over-expression driven by the β-actin promoter reduced cytokine production in DCs following LPS stimulation.Flt3L-cultured BMDCs (A) from both WT and CD2-SRG3 Tg B6 mice and (B) from both WT and β-actin-SRG3 Tg B6 mice were stimulated with either vehicle or LPS (40, 200, or 1000 ng/ml) for 14 hrs, and subsequently, the percentage of the IL12p40-expressing population among the CD11c+ BMDCs was assessed via flow cytometry. Representative data from three independent experiments are shown (upper panel). The graph in lower panel represents mean percentage ± SD for the proportion of IL12p40 (n = 3; **P<0.01). (C) WT, β-actin-SRG3 Tg, and CD2-SRG3 Tg mice were i.p. injected with LPS (2 μg) or vehicle. Sixteen hrs later, intracellular TNFα, IL12p40, iNOS, and IL10 production was assessed in splenic CD11c+ DCs via flow cytometry. The means ± SD are shown in the graphs (n = 4; *P<0.05, **P<0.01).
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Related In: Results  -  Collection

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pone.0132329.g001: SRG3 over-expression driven by the β-actin promoter reduced cytokine production in DCs following LPS stimulation.Flt3L-cultured BMDCs (A) from both WT and CD2-SRG3 Tg B6 mice and (B) from both WT and β-actin-SRG3 Tg B6 mice were stimulated with either vehicle or LPS (40, 200, or 1000 ng/ml) for 14 hrs, and subsequently, the percentage of the IL12p40-expressing population among the CD11c+ BMDCs was assessed via flow cytometry. Representative data from three independent experiments are shown (upper panel). The graph in lower panel represents mean percentage ± SD for the proportion of IL12p40 (n = 3; **P<0.01). (C) WT, β-actin-SRG3 Tg, and CD2-SRG3 Tg mice were i.p. injected with LPS (2 μg) or vehicle. Sixteen hrs later, intracellular TNFα, IL12p40, iNOS, and IL10 production was assessed in splenic CD11c+ DCs via flow cytometry. The means ± SD are shown in the graphs (n = 4; *P<0.05, **P<0.01).
Mentions: DCs play an important role in T cell polarization in the periphery and in the CNS. To determine whether the over-expression of SRG3 has any influence on DC activation elicited by LPS stimulation, BMDCs were generated from CD2-SRG3 Tg, β-actin-SRG3 Tg, and wild type (WT) B6 mice in the presence of Flt3L. After BMDCs were stimulated with LPS, their activation status was monitored based on their IL12p40 cytokine expression pattern. BMDCs from CD2-SRG3 Tg mice produced comparable levels of IL12p40 than those from WT B6 mice (Fig 1A). However, Intriguingly, BMDCs from β-actin-SRG3 Tg mice produced much lower levels of IL12p40 than those from WT B6 mice (Fig 1B), which suggested that the over-expression of SRG3 in DCs restrained the activation of DCs by reducing the production of pro-inflammatory cytokines such as IL12p40. Next, to determine whether SRG3 over-expression affects DC activation by LPS in vivo, β-actin-SRG3 Tg, CD2-SRG3 Tg, and WT B6 mice were i.p. injected with LPS (2 μg) or vehicle. Sixteen hrs later, the expression of TNFα, IL12p40, inducible nitric oxide synthase (iNOS), and IL10 in splenic DCs was examined via flow cytometric analysis. Strikingly, the expression levels of TNFα, IL12p40, and iNOS were decreased by SRG3 over-expression driven by the β-actin promoter. However, as expected, SRG3 over-expression driven by the CD2 promoter did not affect cytokine production in DCs because DCs from CD2-SRG3 Tg mice do not over-express SRG3, as these cells lack CD2 expression. In the vehicle-injected groups, DCs from β-actin-SRG3, but not CD2-SRG3 Tg mice expressed a much lower level of IL12p40 and iNOS but comparable levels of TNFα and IL10 compared to those from WT B6 mice, indicating that SRG3 over-expression in DCs maintains a low basal level of pro-inflammatory gene expression. In addition, no change in IL10 expression was observed between WT B6 mice and either CD2-SRG3 or β-actin-SRG3 Tg mice (Fig 1C; S1 Fig). As expected, the expression levels of SRG3 in splenic CD4+ T cells from both CD2-SRG3 Tg and β-actin-SRG3 Tg mice were up-regulated compared to those from WT B6 mice, whereas the expression levels of SRG3 in splenic DCs were up-regulated in β-actin-SRG3 Tg but not in CD2-SRG3 Tg mice compared to those from WT B6 mice (Fig 1D). These data demonstrated that SRG3 over-expression driven by the β-actin promoter, but not the CD2 promoter, led to the decreased expression of pro-inflammatory cytokines upon LPS stimulation both in vivo and in vitro.

Bottom Line: We found that the outcome of EAE development was completely different depending on the expression pattern of SRG3.SRG3 over-expression not only reduced pro-inflammatory cytokine production by DCs but also shifted macrophages from the inducible nitric oxide synthase (iNOS)-expressing M1 phenotype to the arginase-1-expressing M2 phenotype during EAE.Taken together, our results provide the first evidence supporting the development of a new therapeutic strategy for EAE involving the modulation of SRG3 expression to induce M2 and Th2 polarization, thereby inhibiting inflammatory immune responses.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Bioscience and Biotechnology, Institute of Bioscience, Sejong University, Seoul 143-747, Korea; School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Korea.

ABSTRACT
Although SWI3-related gene (SRG3), a chromatin remodeling factor, is critical for various biological processes including early embryogenesis and thymocyte development, it is unclear whether SRG3 is involved in the differentiation of CD4+ T cells, the key mediator of adaptive immune responses. Because it is known that experimental autoimmune encephalomyelitis (EAE) development is determined by the activation of CD4+ T helper cells, here, we investigated the role of SRG3 in EAE development using SRG3 transgenic mouse models exhibiting two distinct SRG3 expression patterns: SRG3 expression driven by either the CD2 or β-actin promoter. We found that the outcome of EAE development was completely different depending on the expression pattern of SRG3. The specific over-expression of SRG3 using the CD2 promoter facilitated EAE via the induction of Th1 and Th17 cells, whereas the ubiquitous over-expression of SRG3 using the β-actin promoter inhibited EAE by promoting Th2 differentiation and suppressing Th1 and Th17 differentiation. In addition, the ubiquitous over-expression of SRG3 polarized CD4+ T cell differentiation towards the Th2 phenotype by converting dendritic cells (DCs) or macrophages to Th2 types. SRG3 over-expression not only reduced pro-inflammatory cytokine production by DCs but also shifted macrophages from the inducible nitric oxide synthase (iNOS)-expressing M1 phenotype to the arginase-1-expressing M2 phenotype during EAE. In addition, Th2 differentiation in β-actin-SRG3 Tg mice during EAE was associated with an increase in the basophil and mast cell populations and in IL4 production. Furthermore, the increased frequency of Treg cells in the spinal cord of β-actin-SRG3 Tg mice might induce the suppression of and accelerate the recovery from EAE symptoms. Taken together, our results provide the first evidence supporting the development of a new therapeutic strategy for EAE involving the modulation of SRG3 expression to induce M2 and Th2 polarization, thereby inhibiting inflammatory immune responses.

No MeSH data available.


Related in: MedlinePlus