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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

β-actin promoter-driven SRG3 over-expression increases both the basophil and mast cell numbers and IL4 production.(A) Splenocytes were isolated from the spleens of WT, CD2-SRG3 Tg, and β-actin-SRG3 Tg B6 mice at either 8 or 20 weeks of age. The frequencies of both mast cells (FcεRI+CD200R-CD3-B220-) and basophils (FcεRI+CD200R+CD3-B220-) in the spleen were plotted (upper left panel). The absolute cell numbers of both basophils and mast cells were determined (upper right panels). The means ± SD are shown (n = 3–4; **P<0.01, ***P<0.001). (B) Splenocytes from WT, CD2-SRG3 Tg, and β-actin-SRG3 Tg B6 mice at the age of either 8 or 20 weeks were cultured in the presence of recombinant mIL3 (20 ng/ml) for 24 hrs. The percentages of IL4-producing cells among both basophils and mast cells were analyzed via flow cytometry. The mean values ± SD are shown (n = 3–4; *P<0.05, **P<0.01). (c) Splenocytes were isolated from the spleens of WT, CD2-SRG3 Tg, and β-actin-SRG3 Tg B6 mice at either 8 or 20 weeks of age. The absolute cell numbers of both IL4+ basophils and IL4+ mast cells were determined. The means ± SD are shown (n = 3–4; **P<0.01, ***P<0.001).
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pone.0132329.g003: β-actin promoter-driven SRG3 over-expression increases both the basophil and mast cell numbers and IL4 production.(A) Splenocytes were isolated from the spleens of WT, CD2-SRG3 Tg, and β-actin-SRG3 Tg B6 mice at either 8 or 20 weeks of age. The frequencies of both mast cells (FcεRI+CD200R-CD3-B220-) and basophils (FcεRI+CD200R+CD3-B220-) in the spleen were plotted (upper left panel). The absolute cell numbers of both basophils and mast cells were determined (upper right panels). The means ± SD are shown (n = 3–4; **P<0.01, ***P<0.001). (B) Splenocytes from WT, CD2-SRG3 Tg, and β-actin-SRG3 Tg B6 mice at the age of either 8 or 20 weeks were cultured in the presence of recombinant mIL3 (20 ng/ml) for 24 hrs. The percentages of IL4-producing cells among both basophils and mast cells were analyzed via flow cytometry. The mean values ± SD are shown (n = 3–4; *P<0.05, **P<0.01). (c) Splenocytes were isolated from the spleens of WT, CD2-SRG3 Tg, and β-actin-SRG3 Tg B6 mice at either 8 or 20 weeks of age. The absolute cell numbers of both IL4+ basophils and IL4+ mast cells were determined. The means ± SD are shown (n = 3–4; **P<0.01, ***P<0.001).

Mentions: IL4 is a cytokine that induces the M2 polarization of macrophages [11], and basophil-derived IL4 has been reported to represent an effector of the generation of M2 macrophages [27]. Because basophils and mast cells are capable of producing IL4, we examined whether the over-expression of SRG3 modulates the frequencies of basophils and mast cells and the levels of IL4 secretion by these cells in the absence or presence of rIL3 stimulation. Based on the previous study [28], splenic basophils were gated on FcεRI+CD200R+CD3-B220- populations mostly expressing basophil marker DX5 and could be distinguished from splenic mast cells gated on FcεRI+CD200R-CD3-B220- populations expressing mast cell marker c-kit (S3 Fig). We found that even under unstimulated conditions, the over-expression of SRG3 driven by the β-actin promoter, but not the CD2 promoter, elevated the number of basophils and mast cells in the spleen. Moreover, 20-week-old β-actin SRG3 Tg mice displayed a significantly higher number of basophils than 8-week-old mice, whereas the relative increase in the number of basophils from 8 to 20 weeks of age was similar between the β-actin-SRG3 Tg and WT B6 mice. In contrast, the CD2-SRG3 Tg mice exhibited the suppressed development of basophils compared with WT B6 mice at 20, but not 8, weeks of age (Fig 3A). Furthermore, splenic basophils from β-actin-SRG3, but not CD2-SRG3, Tg mice produced a larger amount of IL4 in both the absence and presence of rIL3 stimulation than those from WT B6 mice, and similarly, mast cells showed a tendency towards increased IL4 production under both conditions (Fig 3B). Under unstimulated conditions, the over-expression of SRG3 driven by the β-actin promoter, but not the CD2 promoter, elevated the number of IL4-producing basophils in the spleen at 8 weeks of age, whereas β-actin-SRG3 Tg mice but not CD2-SRG3 Tg mice displayed a significantly higher number of both IL4-producing basophils and IL4-producing mast cells than WT B6 mice at 20-week-old age (Fig 3C). However, both β-actin-SRG3 and CD2-SRG3 Tg mice showed no significant difference in cell number and IL4 production of eosinophils and NKT cells (S4 Fig). These results indicate that the expansion and increased IL4 production of basophils and mast cells were correlated with anti-inflammatory function by SRG3 over-expression.


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)

β-actin promoter-driven SRG3 over-expression increases both the basophil and mast cell numbers and IL4 production.(A) Splenocytes were isolated from the spleens of WT, CD2-SRG3 Tg, and β-actin-SRG3 Tg B6 mice at either 8 or 20 weeks of age. The frequencies of both mast cells (FcεRI+CD200R-CD3-B220-) and basophils (FcεRI+CD200R+CD3-B220-) in the spleen were plotted (upper left panel). The absolute cell numbers of both basophils and mast cells were determined (upper right panels). The means ± SD are shown (n = 3–4; **P<0.01, ***P<0.001). (B) Splenocytes from WT, CD2-SRG3 Tg, and β-actin-SRG3 Tg B6 mice at the age of either 8 or 20 weeks were cultured in the presence of recombinant mIL3 (20 ng/ml) for 24 hrs. The percentages of IL4-producing cells among both basophils and mast cells were analyzed via flow cytometry. The mean values ± SD are shown (n = 3–4; *P<0.05, **P<0.01). (c) Splenocytes were isolated from the spleens of WT, CD2-SRG3 Tg, and β-actin-SRG3 Tg B6 mice at either 8 or 20 weeks of age. The absolute cell numbers of both IL4+ basophils and IL4+ mast cells were determined. The means ± SD are shown (n = 3–4; **P<0.01, ***P<0.001).
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pone.0132329.g003: β-actin promoter-driven SRG3 over-expression increases both the basophil and mast cell numbers and IL4 production.(A) Splenocytes were isolated from the spleens of WT, CD2-SRG3 Tg, and β-actin-SRG3 Tg B6 mice at either 8 or 20 weeks of age. The frequencies of both mast cells (FcεRI+CD200R-CD3-B220-) and basophils (FcεRI+CD200R+CD3-B220-) in the spleen were plotted (upper left panel). The absolute cell numbers of both basophils and mast cells were determined (upper right panels). The means ± SD are shown (n = 3–4; **P<0.01, ***P<0.001). (B) Splenocytes from WT, CD2-SRG3 Tg, and β-actin-SRG3 Tg B6 mice at the age of either 8 or 20 weeks were cultured in the presence of recombinant mIL3 (20 ng/ml) for 24 hrs. The percentages of IL4-producing cells among both basophils and mast cells were analyzed via flow cytometry. The mean values ± SD are shown (n = 3–4; *P<0.05, **P<0.01). (c) Splenocytes were isolated from the spleens of WT, CD2-SRG3 Tg, and β-actin-SRG3 Tg B6 mice at either 8 or 20 weeks of age. The absolute cell numbers of both IL4+ basophils and IL4+ mast cells were determined. The means ± SD are shown (n = 3–4; **P<0.01, ***P<0.001).
Mentions: IL4 is a cytokine that induces the M2 polarization of macrophages [11], and basophil-derived IL4 has been reported to represent an effector of the generation of M2 macrophages [27]. Because basophils and mast cells are capable of producing IL4, we examined whether the over-expression of SRG3 modulates the frequencies of basophils and mast cells and the levels of IL4 secretion by these cells in the absence or presence of rIL3 stimulation. Based on the previous study [28], splenic basophils were gated on FcεRI+CD200R+CD3-B220- populations mostly expressing basophil marker DX5 and could be distinguished from splenic mast cells gated on FcεRI+CD200R-CD3-B220- populations expressing mast cell marker c-kit (S3 Fig). We found that even under unstimulated conditions, the over-expression of SRG3 driven by the β-actin promoter, but not the CD2 promoter, elevated the number of basophils and mast cells in the spleen. Moreover, 20-week-old β-actin SRG3 Tg mice displayed a significantly higher number of basophils than 8-week-old mice, whereas the relative increase in the number of basophils from 8 to 20 weeks of age was similar between the β-actin-SRG3 Tg and WT B6 mice. In contrast, the CD2-SRG3 Tg mice exhibited the suppressed development of basophils compared with WT B6 mice at 20, but not 8, weeks of age (Fig 3A). Furthermore, splenic basophils from β-actin-SRG3, but not CD2-SRG3, Tg mice produced a larger amount of IL4 in both the absence and presence of rIL3 stimulation than those from WT B6 mice, and similarly, mast cells showed a tendency towards increased IL4 production under both conditions (Fig 3B). Under unstimulated conditions, the over-expression of SRG3 driven by the β-actin promoter, but not the CD2 promoter, elevated the number of IL4-producing basophils in the spleen at 8 weeks of age, whereas β-actin-SRG3 Tg mice but not CD2-SRG3 Tg mice displayed a significantly higher number of both IL4-producing basophils and IL4-producing mast cells than WT B6 mice at 20-week-old age (Fig 3C). However, both β-actin-SRG3 and CD2-SRG3 Tg mice showed no significant difference in cell number and IL4 production of eosinophils and NKT cells (S4 Fig). These results indicate that the expansion and increased IL4 production of basophils and mast cells were correlated with anti-inflammatory function by SRG3 over-expression.

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