Influence of sirolimus-induced TGF-β secretion on mouse Treg cell proliferation

J. N. Li, J. X. Li, H. L. Huang, X. L. Zhu, H. Ding, C. F. Huang, J. Lin, J. G. Huang, Z. C. Wu, Muhammad Ashraf, Y. G. Wang, X. K. Li, S. Y. Zheng, J. M. Chen, H. M. Guo, J. Zhuang, P. Zhu

Research output: Contribution to journalArticlepeer-review

8 Scopus citations


We examined the effects of co-culturing CD4+ CD25+ Treg cells with sirolimus or cyclosporin A on Treg cell proliferation and differentiation and on transforming growth factor-β (TGF-β) and Foxp3 expression. CD4+ CD25+ Treg cells were harvested from mononuclear cells of spleens of C57BL/6 mice using immunomagnetic beads and divided into control, sirolimus, and cyclosporine groups. Following a 96-h co-culture, Treg cells were assayed by flow cytometry. FoxP3 and TGF-β mRNA levels and secretion were assayed by reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay. Smad protein of the TGF-β signaling pathway was assayed by western blot and its effect on CD4+ CD25+ FoxP3+ Treg cell proliferation was determined. Sirolimus-promoted differentiation and proliferation was examined using a TGF-β neutralizing antibody. Sirolimus-treated CD4+ T cell TGF-β secretion increased 2.5X over control levels (P < 0.01), but that of the cyclosporine group decreased marginally (P > 0.05). The CD4+ cell proportion decreased significantly (41.25 vs 69.22%, P < 0.01) and slightly (65.21 vs 69.22, P > 0.05) in the cyclosporine and sirolimus groups, respectively. T cell Foxp3 mRNA expression was significantly higher in the sirolimus-treated than in the cyclosporine (53.7 vs 40.2%, P < 0.05) and control groups (P < 0.01), but was significantly lower in the cyclosporine group than in controls (23.6 vs 40.2%, P < 0.01). Overall, sirolimus promoted CD4+ CD25+ Treg cell proliferation and growth in vitro, whereas cyclosporin A inhibited proliferation. Sirolimus might promote CD4+ CD25+ FoxP3+ regulatory T cell proliferation by inducing TGF-β secretion in vivo.

Original languageEnglish (US)
Pages (from-to)18569-18579
Number of pages11
JournalGenetics and Molecular Research
Issue number4
StatePublished - Dec 28 2015
Externally publishedYes


  • Cyclosporin A
  • Foxp3
  • Mice
  • Regulatory T cells
  • Sirolimus
  • Transforming growth factor-β

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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