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Updated: Oct 26, 2025
In Vitro and In Vivo Assessment of T, B and Myeloid Cells Suppressive Activity and Humoral Responses from Transplant Recipients
Published on: August 12, 2017
M-CSF and prostratin induced Mregs promote immune tolerance in transplanted mice through Arg-1 pathway
Jiawei Ji1, Yuan Zhuang2, Haozhou Wang3
1Capital Medical University, Beijing, China; Department of Urology, Capital Medical University Beijing Chaoyang Hospital, Beijing, China.
Objective:
Regulatory macrophages (Mregs) are a group of heterogeneous macrophages. These cells could induce immunosuppressive effects through the expression of immune regulatory molecules and cytokines.
Methods:
The differentiation of Mregs was induced by treating bone marrow cells with M-CSF and prostratin in vitro. The cell-phenotypes and immunosuppressive function were determined by flow cytometry. Rt-PCR was employed to assess the mechanisms of Mregs. Skin grafted mouse model was used for in vivo validation.
Results:
Mregs induced by M-CSF + prostratin had a strong inhibitory effect on T cell proliferation and cytokines production. The phenotype of induced bone marrow cells changed towards Mregs. These Mregs could induce the differentiation of Tregs in vivo. Arg-1 expression in these cells were significantly upregulated. Inhibition of arginase (Arg) or arginine supplement significantly reversed the immunosuppressive function. In mice skin-grafted models, adoptive transfer of these Mregs significantly prolonged allograft survival. In mice models, Arg-1 expression significantly elevated on skin grafts cells and Tregs increased in graft tissues.
Conclusions:
We successfully developed a Mregs-inducing protocol with the combination of M-CSF and prostratin in vitro. M-CSF + prostratin induced Mregs prevented mice skin graft rejection through upregulating the expression Arg-1.
Insights
We developed a new method to create regulatory macrophages (Mregs) using M-CSF and prostratin. These Mregs effectively prevent skin graft rejection in mice by upregulating Arg-1 expression.
Area of Science:
- Immunology
- Cell Biology
- Transplantation Immunology
Background:
- Regulatory macrophages (Mregs) are a heterogeneous cell population known for their immunosuppressive properties.
- Mregs exert immune regulation via specific molecules and cytokines, influencing immune responses.
Purpose of the Study:
- To establish an effective in vitro protocol for inducing regulatory macrophages (Mregs).
- To investigate the immunosuppressive function and mechanisms of Mregs generated using M-CSF and prostratin.
- To validate the efficacy of Mregs in preventing skin graft rejection in vivo.
Main Methods:
- Bone marrow cells were treated with M-CSF and prostratin to induce Mregs in vitro.
- Flow cytometry was used to analyze cell phenotypes and immunosuppressive activity.
- RT-PCR assessed Mreg mechanisms, and skin-grafted mouse models were used for in vivo validation.
Main Results:
- Mregs induced by M-CSF + prostratin demonstrated potent inhibition of T cell proliferation and cytokine production.
- Induced cells exhibited Mreg phenotype, promoted regulatory T cell (Treg) differentiation in vivo, and significantly upregulated Arginase-1 (Arg-1) expression.
- Inhibition of arginase or arginine supplementation reversed immunosuppression; Mregs prolonged allograft survival in mice and increased Tregs in graft tissues.
Conclusions:
- A novel Mregs-inducing protocol utilizing M-CSF and prostratin was successfully developed.
- M-CSF + prostratin-induced Mregs prevent skin graft rejection in mice by upregulating Arg-1 expression.
