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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

The contrasting effects of differential SRG3 over-expression on the severity of EAE were associated with the alteration of the phenotypes of DCs and macrophages.(A) Splenocytes were prepared from both MBP TCR Tg B10.PL mice and CD2-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. The intracellular production of IL6, TNFα, iNOS, and IL10 was assessed in splenic CD11c+ DCs via flow cytometry (upper left panel). Additionally, intracellular IL6, TNFα, iNOS, arginase-1, and IL10 production was assessed in splenic macrophages (CD11c-CD11b+F4/80+) via flow cytometry (upper right panel). Alternatively, splenocytes were prepared from both MBP TCR Tg B10.PL mice and β-actin-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. Intracellular IL6, TNFα, iNOS, and IL10 production was assessed in splenic DCs (CD11c+) (lower left panel) and intracellular IL6, TNFα, iNOS, arginase-1, and IL10 production was assessed in splenic macrophages (CD11c-CD11b+F4/80+) via flow cytometry (lower right panel). The mean values ± SD are shown (n = 5; *P<0.05, **P<0.01). (B) Mononuclear cells (MNCs) were isolated from the spinal cord of both MBP TCR Tg B10.PL mice and CD2-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. The intracellular expression of arginase-1 and iNOS was assessed in macrophages/microglia (CD45+CD11b+F4/80+) via flow cytometry (left panel). Alternatively, MNCs were isolated from the spinal cord of both MBP TCR Tg B10.PL mice and β-actin-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. The intracellular expression of arginase-1 and iNOS was assessed in macrophages/microglia (CD45+CD11b+F4/80+) via flow cytometry (right panel). The mean values ± SD are shown (n = 5; *P<0.05).
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pone.0132329.g006: The contrasting effects of differential SRG3 over-expression on the severity of EAE were associated with the alteration of the phenotypes of DCs and macrophages.(A) Splenocytes were prepared from both MBP TCR Tg B10.PL mice and CD2-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. The intracellular production of IL6, TNFα, iNOS, and IL10 was assessed in splenic CD11c+ DCs via flow cytometry (upper left panel). Additionally, intracellular IL6, TNFα, iNOS, arginase-1, and IL10 production was assessed in splenic macrophages (CD11c-CD11b+F4/80+) via flow cytometry (upper right panel). Alternatively, splenocytes were prepared from both MBP TCR Tg B10.PL mice and β-actin-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. Intracellular IL6, TNFα, iNOS, and IL10 production was assessed in splenic DCs (CD11c+) (lower left panel) and intracellular IL6, TNFα, iNOS, arginase-1, and IL10 production was assessed in splenic macrophages (CD11c-CD11b+F4/80+) via flow cytometry (lower right panel). The mean values ± SD are shown (n = 5; *P<0.05, **P<0.01). (B) Mononuclear cells (MNCs) were isolated from the spinal cord of both MBP TCR Tg B10.PL mice and CD2-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. The intracellular expression of arginase-1 and iNOS was assessed in macrophages/microglia (CD45+CD11b+F4/80+) via flow cytometry (left panel). Alternatively, MNCs were isolated from the spinal cord of both MBP TCR Tg B10.PL mice and β-actin-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. The intracellular expression of arginase-1 and iNOS was assessed in macrophages/microglia (CD45+CD11b+F4/80+) via flow cytometry (right panel). The mean values ± SD are shown (n = 5; *P<0.05).

Mentions: Previously, it has been demonstrated that APCs such as DCs and macrophages play important roles in the pathogenesis of EAE. Specifically, the response and polarization of macrophages and microglia in the CNS are directly involved in the pathogenesis of EAE [12]. Clearly, DCs also participate in the pathological process by presenting self-antigens to CD4+ T cells during EAE. Therefore, we investigated the impact of SRG3 over-expression on DC and macrophage activation during EAE induction. During EAE pathogenesis, CD2 promoter-driven SRG3 over-expression slightly increased the expression of IL6, TNFα, and iNOS, whereas β-actin promoter-driven SRG3 over-expression attenuated the expression of IL6, TNFα, and iNOS. In addition, in the macrophage population, the effect of SRG3 over-expression on macrophage differentiation was remarkable. Regarding the expression of M1 markers (IL6, TNFα, and iNOS) and M2 markers (arginase-1 and IL10), CD2 promoter-driven SRG3 over-expression slightly increased the expression of M1 markers but not M2 markers, whereas β-actin promoter-driven SRG3 over-expression significantly decreased the expression of M1 markers and dramatically increased the expression of M2 markers (Fig 6A). It has been reported that a shift towards the pro-inflammatory M1 phenotype promotes severe EAE but that a shift towards the M2 phenotype suppresses EAE in macrophages and microglia [26, 31]. Thus, we examined whether the effect of SRG3 over-expression on the macrophage differentiation affects macrophages in the spinal cord of EAE-induced mice. As expected, CD2 promoter-driven SRG3 over-expression in EAE-induced mice promoted the M1 phenotype (iNOS) and suppressed the M2 phenotype (arginase-1) in macrophages that infiltrated the spinal cord compared to endogenous SRG3 expression. In contrast, β-actin promoter-driven SRG3 over-expression shifted macrophages from the classical M1 type toward the M2 phenotype in the spinal cord of EAE-induced mice (Fig 6B). Although the numbers of both M1 and M2 macrophages infiltrated into the spinal cord were significantly increased in CD2-SRG3/MBP TCR double Tg mice compared to the control mice, proportion of M1 macrophages was up to two times higher than M2 macrophages. However, the number of M1 but not M2 macrophages infiltrated into the spinal cord was significantly reduced in β-actin-SRG3/MBP TCR double Tg mice compared to the control mice (Fig A in S7 Fig). It has been recently shown that an imbalance of macrophage M1–M2 polarization is often associated with various diseases or inflammatory conditions [32]. M1/M2 ratio among macrophages/microglia from the spinal cord but not the spleen was significantly increased in CD2-SRG3/MBP TCR double Tg mice compared to the control mice, but M1/M2 ratio among those from both the spleen and spinal cord decreased in β-actin-SRG3/MBP TCR double Tg mice compared to the control mice (Fig B in S7 Fig). Thus, our results showed that the contrasting effects between CD2 and β-actin promoter-driven SRG3 over-expression on the severity of EAE were associated with the alteration of the M1/M2 ratio and the inflammatory phenotype of DCs.


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)

The contrasting effects of differential SRG3 over-expression on the severity of EAE were associated with the alteration of the phenotypes of DCs and macrophages.(A) Splenocytes were prepared from both MBP TCR Tg B10.PL mice and CD2-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. The intracellular production of IL6, TNFα, iNOS, and IL10 was assessed in splenic CD11c+ DCs via flow cytometry (upper left panel). Additionally, intracellular IL6, TNFα, iNOS, arginase-1, and IL10 production was assessed in splenic macrophages (CD11c-CD11b+F4/80+) via flow cytometry (upper right panel). Alternatively, splenocytes were prepared from both MBP TCR Tg B10.PL mice and β-actin-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. Intracellular IL6, TNFα, iNOS, and IL10 production was assessed in splenic DCs (CD11c+) (lower left panel) and intracellular IL6, TNFα, iNOS, arginase-1, and IL10 production was assessed in splenic macrophages (CD11c-CD11b+F4/80+) via flow cytometry (lower right panel). The mean values ± SD are shown (n = 5; *P<0.05, **P<0.01). (B) Mononuclear cells (MNCs) were isolated from the spinal cord of both MBP TCR Tg B10.PL mice and CD2-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. The intracellular expression of arginase-1 and iNOS was assessed in macrophages/microglia (CD45+CD11b+F4/80+) via flow cytometry (left panel). Alternatively, MNCs were isolated from the spinal cord of both MBP TCR Tg B10.PL mice and β-actin-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. The intracellular expression of arginase-1 and iNOS was assessed in macrophages/microglia (CD45+CD11b+F4/80+) via flow cytometry (right panel). The mean values ± SD are shown (n = 5; *P<0.05).
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pone.0132329.g006: The contrasting effects of differential SRG3 over-expression on the severity of EAE were associated with the alteration of the phenotypes of DCs and macrophages.(A) Splenocytes were prepared from both MBP TCR Tg B10.PL mice and CD2-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. The intracellular production of IL6, TNFα, iNOS, and IL10 was assessed in splenic CD11c+ DCs via flow cytometry (upper left panel). Additionally, intracellular IL6, TNFα, iNOS, arginase-1, and IL10 production was assessed in splenic macrophages (CD11c-CD11b+F4/80+) via flow cytometry (upper right panel). Alternatively, splenocytes were prepared from both MBP TCR Tg B10.PL mice and β-actin-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. Intracellular IL6, TNFα, iNOS, and IL10 production was assessed in splenic DCs (CD11c+) (lower left panel) and intracellular IL6, TNFα, iNOS, arginase-1, and IL10 production was assessed in splenic macrophages (CD11c-CD11b+F4/80+) via flow cytometry (lower right panel). The mean values ± SD are shown (n = 5; *P<0.05, **P<0.01). (B) Mononuclear cells (MNCs) were isolated from the spinal cord of both MBP TCR Tg B10.PL mice and CD2-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. The intracellular expression of arginase-1 and iNOS was assessed in macrophages/microglia (CD45+CD11b+F4/80+) via flow cytometry (left panel). Alternatively, MNCs were isolated from the spinal cord of both MBP TCR Tg B10.PL mice and β-actin-SRG3/MBP TCR double Tg B10.PL mice immunized with MBP to induce EAE. The intracellular expression of arginase-1 and iNOS was assessed in macrophages/microglia (CD45+CD11b+F4/80+) via flow cytometry (right panel). The mean values ± SD are shown (n = 5; *P<0.05).
Mentions: Previously, it has been demonstrated that APCs such as DCs and macrophages play important roles in the pathogenesis of EAE. Specifically, the response and polarization of macrophages and microglia in the CNS are directly involved in the pathogenesis of EAE [12]. Clearly, DCs also participate in the pathological process by presenting self-antigens to CD4+ T cells during EAE. Therefore, we investigated the impact of SRG3 over-expression on DC and macrophage activation during EAE induction. During EAE pathogenesis, CD2 promoter-driven SRG3 over-expression slightly increased the expression of IL6, TNFα, and iNOS, whereas β-actin promoter-driven SRG3 over-expression attenuated the expression of IL6, TNFα, and iNOS. In addition, in the macrophage population, the effect of SRG3 over-expression on macrophage differentiation was remarkable. Regarding the expression of M1 markers (IL6, TNFα, and iNOS) and M2 markers (arginase-1 and IL10), CD2 promoter-driven SRG3 over-expression slightly increased the expression of M1 markers but not M2 markers, whereas β-actin promoter-driven SRG3 over-expression significantly decreased the expression of M1 markers and dramatically increased the expression of M2 markers (Fig 6A). It has been reported that a shift towards the pro-inflammatory M1 phenotype promotes severe EAE but that a shift towards the M2 phenotype suppresses EAE in macrophages and microglia [26, 31]. Thus, we examined whether the effect of SRG3 over-expression on the macrophage differentiation affects macrophages in the spinal cord of EAE-induced mice. As expected, CD2 promoter-driven SRG3 over-expression in EAE-induced mice promoted the M1 phenotype (iNOS) and suppressed the M2 phenotype (arginase-1) in macrophages that infiltrated the spinal cord compared to endogenous SRG3 expression. In contrast, β-actin promoter-driven SRG3 over-expression shifted macrophages from the classical M1 type toward the M2 phenotype in the spinal cord of EAE-induced mice (Fig 6B). Although the numbers of both M1 and M2 macrophages infiltrated into the spinal cord were significantly increased in CD2-SRG3/MBP TCR double Tg mice compared to the control mice, proportion of M1 macrophages was up to two times higher than M2 macrophages. However, the number of M1 but not M2 macrophages infiltrated into the spinal cord was significantly reduced in β-actin-SRG3/MBP TCR double Tg mice compared to the control mice (Fig A in S7 Fig). It has been recently shown that an imbalance of macrophage M1–M2 polarization is often associated with various diseases or inflammatory conditions [32]. M1/M2 ratio among macrophages/microglia from the spinal cord but not the spleen was significantly increased in CD2-SRG3/MBP TCR double Tg mice compared to the control mice, but M1/M2 ratio among those from both the spleen and spinal cord decreased in β-actin-SRG3/MBP TCR double Tg mice compared to the control mice (Fig B in S7 Fig). Thus, our results showed that the contrasting effects between CD2 and β-actin promoter-driven SRG3 over-expression on the severity of EAE were associated with the alteration of the M1/M2 ratio and the inflammatory phenotype of DCs.

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