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Glycogen synthase kinase 3β inhibition promotes human iTreg differentiation and suppressive function

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Abstract

Induced regulatory T cells (iTregs) are essential to maintain immunological tolerance, immune homeostasis and prevention of autoimmunity. Some studies suggest that glycogen synthase kinase 3β (GSK3β) is involved in the mouse iTreg differentiation; however, whether GSK3β inhibits or enhances iTreg differentiation is still a matter of controversy. To address this issue, we have utilized human naïve CD4+ T cells and investigated whether GSK3 activity changes during iTreg differentiation and whether altering GSK3 activity influences the development of iTregs and its suppressive function. As a constitutively activated kinase, during iTreg differentiation GSK3β became quickly deactivated (phosphorylated at serine 9), which is dependent on MAPK pathway rather than PI3-kinase/Akt pathway. Our results indicated that inhibition of GSK3β by specific inhibitors, SB216763 or TDZD-8, promoted the differentiation of iTreg and increased their suppressive activity. In contrast, overexpression of GSK3β significantly inhibited iTreg differentiation. Furthermore, GSK3β inhibition enhanced iTreg differentiation through the TGF-β/Smad3 pathway. Taken together, this study demonstrates that inhibition of GSK3β enhances human iTreg differentiation and its suppressive activity, and provides a rationale to target GSK3β as a novel immunotherapeutic strategy.

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Acknowledgments

We thank Prof. Li M (Zhongshan School of Medicine, China) for providing the human GSK3β plasmid. This work was supported by grants from the National Natural Science Foundation of China (No. 81373165 and No. 81273261) and Six Talents Peak in Jiangsu Province of China (2013-WSW-022).

Conflict of interest

The authors declare no financial or commercial conflict of interest.

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Authors and Affiliations

  1. Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, 300 GuangZhou Road, Nanjing, Jiangsu Province, People’s Republic of China

    Yongxiang Xia, Han Zhuo, Yunjie Lu, Lei Deng, Runqiu Jiang, Long Zhang, Qin Zhu, Liyong Pu, Xuehao Wang & Ling Lu

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  1. Yongxiang Xia
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  2. Han Zhuo
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Correspondence to Qin Zhu, Liyong Pu or Xuehao Wang.

Electronic supplementary material

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12026_2015_8635_MOESM1_ESM.jpg

Supplementary material 1 (JPEG 270 kb): Human CD4+ T cells were cultured in medium supplemented with SB216763 or TDZD-8 for 1 hour. Phospho-glycogen synthase(p-GS), GS, GSK3β, and GAPDH protein levels were analyzed by immunoblotting

12026_2015_8635_MOESM2_ESM.jpg

Supplementary material 2 (JPEG 104 kb): Human CD4+ T cells were cultured in medium supplemented with SIS3 in the presence or absence of SB216763 for 1 hour. P-Smad3, Smad3 and GAPDH protein levels were analyzed by immunoblotting

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Xia, Y., Zhuo, H., Lu, Y. et al. Glycogen synthase kinase 3β inhibition promotes human iTreg differentiation and suppressive function. Immunol Res 62, 60–70 (2015). https://doi.org/10.1007/s12026-015-8635-3

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