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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

Effects of SM934 on the polarization of CD4+ T cells in EAE mice.Splenocytes from immunized mice treated with vehicle or SM934 were isolated at the peak of disease (day 18 p.i.) and analyzed. (A) The percentage of Th1, Th17 and Treg cells in the CD4+ gate were analyzed by flow cytometry, values in the bar graphs are the mean ± SEM (n = 4). (B) Splenocytes were re-stimulated with IL-12 (20 ng/ml) or IL-23 (20 ng/ml) for 72 hours, and supernatants were collected to measure IFN-γ and IL-17 levels, respectively. (C) RNA from splenocytes was analyzed by real-time PCR for expression of T-bet, RORγt and Foxp3. 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-g003: Effects of SM934 on the polarization of CD4+ T cells in EAE mice.Splenocytes from immunized mice treated with vehicle or SM934 were isolated at the peak of disease (day 18 p.i.) and analyzed. (A) The percentage of Th1, Th17 and Treg cells in the CD4+ gate were analyzed by flow cytometry, values in the bar graphs are the mean ± SEM (n = 4). (B) Splenocytes were re-stimulated with IL-12 (20 ng/ml) or IL-23 (20 ng/ml) for 72 hours, and supernatants were collected to measure IFN-γ and IL-17 levels, respectively. (C) RNA from splenocytes was analyzed by real-time PCR for expression of T-bet, RORγt and Foxp3. Results were expressed as mean ± SEM. *, p<0.05 compared with vehicle control. Three independent experiments were performed with similar results.

Mentions: To further investigate the possible role of SM934 in helper T cell polarization, we analyzed the CD4+ T cell subsets in spleens from EAE mice at the peak of disease. In accordance with ex vivo results, there was a significant decrease of the percentage of IFN-γ+ producing CD4+ T cell (Th1) and IL-17+ producing CD4+ T cell (Th17), but an increase of the percentage of CD4+CD25+Foxp3+ T cells (Treg) in splenocytes (Figure 3A).


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)

Effects of SM934 on the polarization of CD4+ T cells in EAE mice.Splenocytes from immunized mice treated with vehicle or SM934 were isolated at the peak of disease (day 18 p.i.) and analyzed. (A) The percentage of Th1, Th17 and Treg cells in the CD4+ gate were analyzed by flow cytometry, values in the bar graphs are the mean ± SEM (n = 4). (B) Splenocytes were re-stimulated with IL-12 (20 ng/ml) or IL-23 (20 ng/ml) for 72 hours, and supernatants were collected to measure IFN-γ and IL-17 levels, respectively. (C) RNA from splenocytes was analyzed by real-time PCR for expression of T-bet, RORγt and Foxp3. Results were expressed as mean ± SEM. *, p<0.05 compared with vehicle control. Three independent experiments were performed with similar results.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0074108-g003: Effects of SM934 on the polarization of CD4+ T cells in EAE mice.Splenocytes from immunized mice treated with vehicle or SM934 were isolated at the peak of disease (day 18 p.i.) and analyzed. (A) The percentage of Th1, Th17 and Treg cells in the CD4+ gate were analyzed by flow cytometry, values in the bar graphs are the mean ± SEM (n = 4). (B) Splenocytes were re-stimulated with IL-12 (20 ng/ml) or IL-23 (20 ng/ml) for 72 hours, and supernatants were collected to measure IFN-γ and IL-17 levels, respectively. (C) RNA from splenocytes was analyzed by real-time PCR for expression of T-bet, RORγt and Foxp3. Results were expressed as mean ± SEM. *, p<0.05 compared with vehicle control. Three independent experiments were performed with similar results.
Mentions: To further investigate the possible role of SM934 in helper T cell polarization, we analyzed the CD4+ T cell subsets in spleens from EAE mice at the peak of disease. In accordance with ex vivo results, there was a significant decrease of the percentage of IFN-γ+ producing CD4+ T cell (Th1) and IL-17+ producing CD4+ T cell (Th17), but an increase of the percentage of CD4+CD25+Foxp3+ T cells (Treg) in splenocytes (Figure 3A).

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