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

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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.

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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.

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    Authors and Affiliations

    1. Transplantation Biology Research Division, State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of Sciences, Beichen West Road 1-5, Chaoyang District, Beijing, 100101, China

      Fan Yang, Yang Li, Tingting Wu, Yang Zhao & Yong Zhao

    2. Department of Kidney Transplantation, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong, China

      Ning Na

    3. College of Life Science, Henan Normal University, Xinxiang, Henan, China

      Weiguo Li & Chenlu Han

    4. Key Laboratory of Human Diseases Comparative Medicine, Ministry of Health; Institute of Laboratory Animal Science, CAMS & PUMC, Beijing, China

      Lianfeng Zhang

    5. Hepatology and Cancer Biotherapy Ward, Beijing YouAn Hospital, Capital Medical University, Beijing, 100069, China

      Jun Lu

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    Fan Yang, Yang Li and Tingting Wu contributed equally to this work.

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    Yang, F., Li, Y., Wu, T. et al. TNFα-induced M-MDSCs promote transplant immune tolerance via nitric oxide. J Mol Med 94, 911–920 (2016). https://doi.org/10.1007/s00109-016-1398-z

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