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Artemisinin analogue SM934 ameliorates murine experimental autoimmune encephalomyelitis through enhancing the expansion and functions of regulatory T cell.

Li X, Li TT, Zhang XH, Hou LF, Yang XQ, Zhu FH, Tang W, Zuo JP - PLoS ONE (2013)

Bottom Line: In vivo, administration of SM934 significantly inhibited the development of EAE and suppressed the elevation of serum IL-17.Ex vivo, upon antigen-recall stimulation, IL-2, IFN-γ, IL-17 and IL-6 production were decreased, whereas IL-10 and TGF-β production were increased from the splenocytes isolated from SM934-treated mice.Furthermore, in the spinal lesion, SM934 treatment dramatically decreased the infiltration of CD4(+) T cells, within which the Treg cells percentage was enlarged, whereas the Th17, but not Th1 percentage, was significantly decreased comparing with the vehicle-treated groups.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunopharmacology, State Key Laboratory of Drug Research Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, PR China.

ABSTRACT

Background: Artemisinin analogue SM934 was previously reported to possess immunosuppressive properties. The aim of this study was to determine the effects and the underlying mechanisms of SM934 in murine experimental autoimmune encephalomyelitis (EAE).

Methods: Female C57BL/6 mice immunized with MOG35-55 were treated with or without SM934, then the clinical scores and other relevant parameters were assessed. Th1, Th17 and regulatory T (Treg) cell profiles were determined through ELISA, qRT-PCR, flow cytometry and BrdU incorporation assay. The effects of SM934 on Th1, Th17 and Treg cells differentiation were explored through intracellular staining and flow cytometry examination.

Results: In vivo, administration of SM934 significantly inhibited the development of EAE and suppressed the elevation of serum IL-17. Ex vivo, upon antigen-recall stimulation, IL-2, IFN-γ, IL-17 and IL-6 production were decreased, whereas IL-10 and TGF-β production were increased from the splenocytes isolated from SM934-treated mice. Consistently, both flow cytometry and qRT-PCR results showed that SM934 treatment significantly increased the Treg, while strongly suppressed the Th17 and Th1, responses in the peripheral. Furthermore, in the spinal lesion, SM934 treatment dramatically decreased the infiltration of CD4(+) T cells, within which the Treg cells percentage was enlarged, whereas the Th17, but not Th1 percentage, was significantly decreased comparing with the vehicle-treated groups. Finally, both BrdU incorporation and in vitro Treg differentiation assays revealed that SM934 treatment could directly promote the expansion of Treg cells in vivo and in vitro.

Conclusion: Taken together, this study demonstrated that SM934 treatment could ameliorate the murine EAE disease, which might be mediated by inducing Treg differentiation and expansion.

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Related in: MedlinePlus

SM934 treatment enhanced Treg cells expansion in EAE mice.(A) Vehicle and SM934 treated mice were given daily i.p. injection of BrdU for 3 days from day 17 p.i., splenocytes and MNC in spinal cord were isolated and analyzed for BrdU incorporation. The proliferative profile was evaluated in CD4+Foxp3−, CD4+Foxp3+ splenocytes, and CD4+Foxp3−, CD4+Foxp3+ from spinal cord. (B) Percentage of CD4+Foxp3−, CD4+Foxp3+ splenocytes, and CD4+Foxp3−, CD4+Foxp3+ from spinal cord are expressed as mean ± SEM (n = 4). (C) CD4+ T cells were purified by immunomagnetic negative selection from splenocytes, indicated genes expression was analyzed by real-time PCR. Results were expressed as mean ± SEM. *, p<0.05 compared with vehicle control. Three independent experiments were performed with similar results.
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pone-0074108-g005: SM934 treatment enhanced Treg cells expansion in EAE mice.(A) Vehicle and SM934 treated mice were given daily i.p. injection of BrdU for 3 days from day 17 p.i., splenocytes and MNC in spinal cord were isolated and analyzed for BrdU incorporation. The proliferative profile was evaluated in CD4+Foxp3−, CD4+Foxp3+ splenocytes, and CD4+Foxp3−, CD4+Foxp3+ from spinal cord. (B) Percentage of CD4+Foxp3−, CD4+Foxp3+ splenocytes, and CD4+Foxp3−, CD4+Foxp3+ from spinal cord are expressed as mean ± SEM (n = 4). (C) CD4+ T cells were purified by immunomagnetic negative selection from splenocytes, indicated genes expression was analyzed by real-time PCR. Results were expressed as mean ± SEM. *, p<0.05 compared with vehicle control. Three independent experiments were performed with similar results.

Mentions: To further determine whether SM934 directly influence the Treg cells in vivo, BrdU incorporation assay was performed in EAE mice and the proliferation of Teff and Treg cells was compared. Results presented in Figure 5A showed that SM934 significantly enhanced Treg cells (CD4+Foxp3+) proliferation both in the peripheral and CNS, while showing no effect on the proliferation of Teff (CD4+Foxp3−).


Artemisinin analogue SM934 ameliorates murine experimental autoimmune encephalomyelitis through enhancing the expansion and functions of regulatory T cell.

Li X, Li TT, Zhang XH, Hou LF, Yang XQ, Zhu FH, Tang W, Zuo JP - PLoS ONE (2013)

SM934 treatment enhanced Treg cells expansion in EAE mice.(A) Vehicle and SM934 treated mice were given daily i.p. injection of BrdU for 3 days from day 17 p.i., splenocytes and MNC in spinal cord were isolated and analyzed for BrdU incorporation. The proliferative profile was evaluated in CD4+Foxp3−, CD4+Foxp3+ splenocytes, and CD4+Foxp3−, CD4+Foxp3+ from spinal cord. (B) Percentage of CD4+Foxp3−, CD4+Foxp3+ splenocytes, and CD4+Foxp3−, CD4+Foxp3+ from spinal cord are expressed as mean ± SEM (n = 4). (C) CD4+ T cells were purified by immunomagnetic negative selection from splenocytes, indicated genes expression was analyzed by real-time PCR. Results were expressed as mean ± SEM. *, p<0.05 compared with vehicle control. Three independent experiments were performed with similar results.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3756992&req=5

pone-0074108-g005: SM934 treatment enhanced Treg cells expansion in EAE mice.(A) Vehicle and SM934 treated mice were given daily i.p. injection of BrdU for 3 days from day 17 p.i., splenocytes and MNC in spinal cord were isolated and analyzed for BrdU incorporation. The proliferative profile was evaluated in CD4+Foxp3−, CD4+Foxp3+ splenocytes, and CD4+Foxp3−, CD4+Foxp3+ from spinal cord. (B) Percentage of CD4+Foxp3−, CD4+Foxp3+ splenocytes, and CD4+Foxp3−, CD4+Foxp3+ from spinal cord are expressed as mean ± SEM (n = 4). (C) CD4+ T cells were purified by immunomagnetic negative selection from splenocytes, indicated genes expression was analyzed by real-time PCR. Results were expressed as mean ± SEM. *, p<0.05 compared with vehicle control. Three independent experiments were performed with similar results.
Mentions: To further determine whether SM934 directly influence the Treg cells in vivo, BrdU incorporation assay was performed in EAE mice and the proliferation of Teff and Treg cells was compared. Results presented in Figure 5A showed that SM934 significantly enhanced Treg cells (CD4+Foxp3+) proliferation both in the peripheral and CNS, while showing no effect on the proliferation of Teff (CD4+Foxp3−).

Bottom Line: In vivo, administration of SM934 significantly inhibited the development of EAE and suppressed the elevation of serum IL-17.Ex vivo, upon antigen-recall stimulation, IL-2, IFN-γ, IL-17 and IL-6 production were decreased, whereas IL-10 and TGF-β production were increased from the splenocytes isolated from SM934-treated mice.Furthermore, in the spinal lesion, SM934 treatment dramatically decreased the infiltration of CD4(+) T cells, within which the Treg cells percentage was enlarged, whereas the Th17, but not Th1 percentage, was significantly decreased comparing with the vehicle-treated groups.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunopharmacology, State Key Laboratory of Drug Research Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, PR China.

ABSTRACT

Background: Artemisinin analogue SM934 was previously reported to possess immunosuppressive properties. The aim of this study was to determine the effects and the underlying mechanisms of SM934 in murine experimental autoimmune encephalomyelitis (EAE).

Methods: Female C57BL/6 mice immunized with MOG35-55 were treated with or without SM934, then the clinical scores and other relevant parameters were assessed. Th1, Th17 and regulatory T (Treg) cell profiles were determined through ELISA, qRT-PCR, flow cytometry and BrdU incorporation assay. The effects of SM934 on Th1, Th17 and Treg cells differentiation were explored through intracellular staining and flow cytometry examination.

Results: In vivo, administration of SM934 significantly inhibited the development of EAE and suppressed the elevation of serum IL-17. Ex vivo, upon antigen-recall stimulation, IL-2, IFN-γ, IL-17 and IL-6 production were decreased, whereas IL-10 and TGF-β production were increased from the splenocytes isolated from SM934-treated mice. Consistently, both flow cytometry and qRT-PCR results showed that SM934 treatment significantly increased the Treg, while strongly suppressed the Th17 and Th1, responses in the peripheral. Furthermore, in the spinal lesion, SM934 treatment dramatically decreased the infiltration of CD4(+) T cells, within which the Treg cells percentage was enlarged, whereas the Th17, but not Th1 percentage, was significantly decreased comparing with the vehicle-treated groups. Finally, both BrdU incorporation and in vitro Treg differentiation assays revealed that SM934 treatment could directly promote the expansion of Treg cells in vivo and in vitro.

Conclusion: Taken together, this study demonstrated that SM934 treatment could ameliorate the murine EAE disease, which might be mediated by inducing Treg differentiation and expansion.

Show MeSH
Related in: MedlinePlus