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Insufficient secretion of IL-10 by Tregs compromised its control on over-activated CD4+ T effector cells in newly diagnosed adult immune thrombocytopenia patients

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Abstract

Primary immune thrombocytopenia (ITP) is an autoimmune heterogeneous disorder. Excessive activated CD4+ T effector cells (Teffs) and compromised regulatory T cells (Tregs) were reported in ITP patients, yet little is known about the mechanisms. Interleukin-10 (IL-10) is an important regulatory cytokine of Tregs in inflammatory condition. It has been recently highlighted that IL-10-producing Tregs contribute to the effective controls of several autoimmune diseases. Hence this study was aimed to examine the role of IL-10 produced by Tregs in newly diagnosed ITP patients. Newly diagnosed ITP patients and healthy volunteers were enrolled to assess the numbers of peripheral Th1, Th17 cells and Tregs. CD4+CD25−Teffs and CD4+CD25+Tregs were purified. CD4+CD25−Teffs, labeled with CFSE, were cultured alone or with Tregs for 5 days, and the supernatants were then collected for IL-10 concentration test. The role of IL-10 in Tregs’ inhibitory function was also determined. In ITP patients, Teffs were excessively activated, while the Tregs were numerically and functionally impaired. The percentages of IL-10+ Tregs in Tregs’ population were found elevated dramatically in ITP patients but decreased in the remitted patients. The IL-10 concentrations in the cultured supernatant were decreased in ITP patients but elevated in the remitted patients. Furthermore, the IL-10 secretion by Tregs was dramatically decreased in ITP patients. IL-10 treatment enhanced the suppression effect of Tregs toward Teffs, whereas anti-IL-10 treatment boosted the proliferation of Teffs and Th17 cells. Excessive activated Teffs and impaired Tregs play major roles in the exuberant CD4+ T cells immune responses of ITP. The inhibitory effect of Tregs toward Teffs is largely exerted by IL-10. Insufficient secretion of IL-10 compromises the inhibitory capability of Tregs against Teffs in newly diagnosed ITP patients.

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Acknowledgments

This work was supported by grants from National Natural Science Foundation of China (30972737, 81170473 and 81300381) and Shanghai Health Bureau Funding (20134y117 and 20124223). All authors obtain permission to acknowledge from all those mentioned in the Acknowledgements.

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The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Department of Hematology, Zhongshan Hospital, Fudan University, 180 Fenglin Rd, Shanghai, 200032, China

    Feng Li, Lili Ji, Weiguang Wang, Fanli Hua, Yanxia Zhan, Shanhua Zou, Ling Yuan, Yang Ke & Yunfeng Cheng

  2. Department of Hematology, Jinshan Hospital, Fudan University, Shanghai, China

    Fanli Hua

  3. Biomedical Research Center, Zhongshan Hospital, Fudan University, Shanghai, China

    Zhihui Min, Dongli Song & Yunfeng Cheng

  4. Department of Hematology, Zhongshan Hospital Qingpu Branch, Shanghai, China

    Lihua Sun & Yunfeng Cheng

  5. Department of Cardiothoracic Surgery, Tongji Hospital, Tongji University, 389 Xincun Rd, Shanghai, 200065, China

    Hao Chen

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  1. Feng Li
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Correspondence to Hao Chen or Yunfeng Cheng.

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Feng Li and Lili Ji have contributed equally to this work as the first authorship.

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12026_2014_8620_MOESM1_ESM.tif

The purification of Tregs and Teffs. Representative dot plots of CD4+T cells, CD4+CD25+ Tregs, and CD4+CD25− Teffs. The purification of each cell type was above 90 % (TIFF 1201 kb)

12026_2014_8620_MOESM2_ESM.tif

Comparative percentage of IL-10+ Teffs was detected in the NC and ITP groups. NC: normal control; ITP: ITP group; p values were indicated in the figure (TIFF 269 kb)

12026_2014_8620_MOESM3_ESM.tif

Isotype control FCM figure for Fig. 1j. It is the isotype control figure for Foxp3 and IL-10 antigen in PBMCs stimulated with PMA, ionomycin and BFA (TIFF 353 kb)

12026_2014_8620_MOESM4_ESM.tif

Isotype control FCM figure for Fig. 2a and c. It is the isotype control figure for IFN-γ and il-17A antigen of CD4+T cells cultured for 5 days, stimulated with PMA, ionomycin and BFA (TIFF 385 kb)

12026_2014_8620_MOESM5_ESM.tif

Isotype control FCM figure for Fig. 3a. It is the isotype control figure for Foxp3 and IL-10 antigen in CD4+CD25+T cells cultured for 5 days, stimulated with PMA, ionomycin and BFA (TIFF 375 kb)

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Li, F., Ji, L., Wang, W. et al. Insufficient secretion of IL-10 by Tregs compromised its control on over-activated CD4+ T effector cells in newly diagnosed adult immune thrombocytopenia patients. Immunol Res 61, 269–280 (2015). https://doi.org/10.1007/s12026-014-8620-2

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