Journal of Molecular Medicine

, Volume 94, Issue 8, pp 911–920 | Cite as

TNFα-induced M-MDSCs promote transplant immune tolerance via nitric oxide

  • Fan Yang
  • Yang Li
  • Tingting Wu
  • Ning Na
  • Yang Zhao
  • Weiguo Li
  • Chenlu Han
  • Lianfeng Zhang
  • Jun Lu
  • Yong Zhao
Original Article
First Online:
Received:
Revised:
Accepted:

Abstract

Efficient induction of functional competent myeloid-derived suppressor cells (MDSCs) will be critical for the clinical application of MDSCs to treat autoimmune diseases and to induce transplantation immune tolerance. In the present study, we tried to establish the MDSC induction system with M-CSF and tumor necrosis factor α (TNFα) and investigated the immunosuppressive function of M-CSF + TNFα-induced MDSCs in transplant mouse models. Monocytic MDSCs (M-MDSCs) were induced by culture of the non-adherent mouse bone marrow cells with M-CSF or M-CSF + TNFα, respectively, for 7 days. Phenotype analysis revealed that the majority of M-CSF- and M-CSF + TNFα-induced MDSCs express F4/80. The addition of TNFα in the induction period increased Gr-1, Ly6C, CD80, and CD274 expressions on these cells. M-CSF + TNFα-induced M-MDSCs showed poor TNFα, IL-12, and IL-6 expressions after lipopolysaccharide (LPS) stimulation and decreased arginase 1 (Arg-1) and Fizz expressions after IL-4 stimulation compared with M-CSF-induced M-MDSCs. M-CSF + TNFα-induced M-MDSCs showed enhanced ability to suppress T cell proliferation and cytokine production than M-CSF-induced M-MDSCs. M-CSF + TNFα-induced M-MDSCs express high levels of inducing nitric oxide synthase (iNOS) and blocking iNOS activity by a chemical inhibitor or gene deficiency significantly reversed the inhibitory effects of M-CSF + TNFα-induced M-MDSCs on T cells. Adoptive transfer of M-CSF + TNFα-induced M-MDSCs promoted immune tolerance in a male-to-female skin-grafted mice, but M-CSF + TNFα-induced iNOS-deficient M-MDSCs failed to do so. Thus, M-CSF + TNFα-induced M-MDSCs have powerful immunosuppressive activity, which is mediated by an iNOS-dependent pathway. M-CSF + TNFα-induced M-MDSCs can promote immune tolerance to donor antigens in a transplant mouse model.

Key message

  • The combination of M-CSF and TNFα efficiently induces functional M-MDSCs in vitro.

  • M-CSF + TNFα-induced M-MDSCs promote immune tolerance in a transplant mouse model.

  • The immunosuppressive ability of M-CSF + TNFα-induced M-MDSCs is dependent on iNOS.

Keywords

Myeloid-derived suppressor cells Immune tolerance Nitric oxide Transplantation 
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Notes

Acknowledgments

The authors wish to thank Dr. Yuzhu Hou a for his kind review of the manuscript; Mrs. Qing Meng, Mr. Yabing Liu, and Dr. Xiaoqiu Liu for their expert technical assistance; Mrs. Ling Li for her excellent laboratory management; and Mr. Baisheng Ren for his outstanding animal husbandry. This work was supported by grants from the National Science and Technology Major Project “prevention and treatment of AIDS and virus hepatitis”(2014ZX10002002-001-002, J.L., Y.Z.), the National Basic Research Program of China (2010CB945301, 2011CB710903, Y.Z.), the National Natural Science Foundation of China for General and Key Programs (C81130055, C81072396, Y.Z.), Knowledge Innovation Program of the Chinese Academy of Sciences (XDA04020202-19, Y.Z.), and the CAS/SAFEA International Partnership Program for Creative Research Teams (Y.Z.).

Author contributions

Fan Yang (hopscotch@126.com), Yang Li (liyang.zhe.happy@163.com), and Tingting Wu (wtt1983_18@aliyun.com) designed and did the experiments, analyzed the data, and contributed to writing the manuscript.

Ning Na (nngg20102009@hotmail.com) analyzed data, comments on the project, and revised the manuscript.

Yang Zhao (dongdong_1221@126.com) performed the flow cytometry assays and analyzed the data.

Weiguo Li (041011@htu.edu.cn) analyzed the data and designed the experiments.

Chenlu Han (hanchenlu110@163.com) did the PCR assays.

Lianfeng Zhang (zhanglf@cnilas.org) provided the animal models and revised the manuscript.

Jun Lu (lujun98@ccmu.edu.cn) and Yong Zhao (zhaoy@ioz.ac.cn) designed the experiments, analyzed the data, wrote the manuscript, and provided the overall direction.

Compliance with ethical standards

Conflict of interest

The authors declare no competing financial interests.

Supplementary material

109_2016_1398_MOESM1_ESM.pdf (490 kb)
ESM 1 (PDF 490 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Fan Yang
    • 1
  • Yang Li
    • 1
  • Tingting Wu
    • 1
  • Ning Na
    • 2
  • Yang Zhao
    • 1
  • Weiguo Li
    • 3
  • Chenlu Han
    • 3
  • Lianfeng Zhang
    • 4
  • Jun Lu
    • 5
  • Yong Zhao
    • 1
  1. 1.Transplantation Biology Research Division, State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of ZoologyChinese Academy of SciencesBeijingChina
  2. 2.Department of Kidney TransplantationThe Third Affiliated Hospital of Sun Yat-sen UniversityGuangzhouChina
  3. 3.College of Life ScienceHenan Normal UniversityXinxiangChina
  4. 4.Key Laboratory of Human Diseases Comparative Medicine, Ministry of Health; Institute of Laboratory Animal ScienceCAMS & PUMCBeijingChina
  5. 5.Hepatology and Cancer Biotherapy Ward, Beijing YouAn HospitalCapital Medical UniversityBeijingChina

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