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CD4+CD25+ regulatory T cells are not required for mesenchymal stem cell function in fully MHC-mismatched mouse cardiac transplantation

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Abstract

Although the immunomodulative properties of mesenchymal stem cells (MSCs) open up attractive possibilities in solid-organ transplantation, information concerning the optimal dose, route, timing of administration, major histocompatibility complex (MHC)-restriction and relevant mechanisms is currently lacking. Therefore, better characterization of MSC immunoregulatory activity and elucidation of its mechanisms are crucial. In this study, we confirmed that MSCs did not elicit proliferation by allogeneic CD4+ T cells, suggesting that MSCs were not immunogenic. By using C57BL/6 mouse MSCs as donor-derived or recipient-derived or as third-party MSCs, we discovered that MSCs suppressed CD4+ T cell proliferation and prolonged mouse cardiac allograft survival in a dose-dependent and non-MHC-restricted manner. We also found that intraperitoneal administration favored survival prolongation, although this prolongation was weaker than that via the intravenous route. Only infusion at earlier time points favored survival prolongation. Depletion of CD4+CD25+ T cells did not affect the immunosuppression of MSCs on CD4+ T cells. Moreover, MSCs did not induce regulatory T cells. The in vivo data revealed that MSCs did not increase the percentage of CD4+CD25+ T cells and FoxP3 expression. More importantly, we demonstrated for the first time that depletion of CD4+CD25+ T cells did not hinder MSC-induced survival prolongation, indicating that CD4+CD25+ regulatory T cells were not essential for the prolongation of MSC-mediated allograft survival.

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Abbreviations

BW:

Bone marrow

CTL:

Cytotoxic T lymphocyte

FACS:

Fluorescence-activated cell sorter

FCS:

Fetal calf serum

FoxP3:

Forkhead/winged helix transcription factor

i.p.:

Intraperitoneally

i.v.:

Intravenously

GVHD:

Graft versus host disease

mAb:

Monoclonal antibody

MACS:

Magnetic-activated cell sorting

MHC:

Major histocompatibility complex

MLR:

Mixed lymphocyte reaction

MSCs:

Mesenchymal stem cells

NK:

Natural killer

PBS:

Phosphate-buffered saline

RT-PCR:

Reverse transcriptase polymerase chain reaction

SD:

Standard deviation

Treg:

CD4+CD25+ regulatory T cells

WT:

Wild-type

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Funding

This work was supported by grants from the Project Sponsored by the Scientific Research Foundation for Returned Overseas Chinese Scholars, State Education Ministry (Project-sponsored by SRF for ROCS, SEM), China (no. 2008890) and from The Educational Department of Liaoning Province, China (no. 2008824). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Conflict of interest

The authors declare no competing interests.

Author Contributions

The experiments were conceived and designed by X.J. and C.L. The experiments were performed by J.H., D.G., J.G., J.Y., K.J., Z.C., L.Z. and L.L. The data were analyzed by J.H., W.S. and J.Y. Reagents/materials/analysis tools were contributed by D.G. and J.L. The manuscript was written by X.J., J.H. and D.G.

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Authors and Affiliations

  1. Department of Surgery, The Fourth Affiliated Hospital, China Medical University, Shenyang, 110032, Liaoning Province, People’s Republic of China

    Xiaofeng Jiang, Jianpeng Hao, Dawei Guo, Jinshuai Guo, Junchao Yao, Kun Jiang, Zheming Cui, Lei Zhu, Wenyu Sun, Lin Lin & Jian Liang

  2. Department of Biliary Surgery, Eastern Hepatobiliary Surgery Hospital, The Second Military Medical University, 200438, Shanghai, People’s Republic of China

    Chen Liu

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  1. Xiaofeng Jiang
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  2. Chen Liu
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Correspondence to Xiaofeng Jiang.

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Jiang, X., Liu, C., Hao, J. et al. CD4+CD25+ regulatory T cells are not required for mesenchymal stem cell function in fully MHC-mismatched mouse cardiac transplantation. Cell Tissue Res 358, 503–514 (2014). https://doi.org/10.1007/s00441-014-1956-z

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