In-vitro immunomodulatory efficacy of extracellular vesicles derived from TGF-β1/IFN-γ dual licensed human bone marrow mesenchymal stromal cells
Abstract Background Mesenchymal stromal cells (MSCs) possess strong immunomodulatory properties, making them attractive candidates for regenerative medicine and immune-related therapies. Pre-activation, or licensing, of MSCs with cytokines such as interferon-gamma (IFN-γ) and transforming growth fac...
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BMC
2025-07-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | https://doi.org/10.1186/s13287-025-04476-2 |
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| author | Jiemin Wang Seyedmohammad Moosavizadeh Manon Jammes Abbas Tabasi Trung Bach Aideen E. Ryan Thomas Ritter |
| author_facet | Jiemin Wang Seyedmohammad Moosavizadeh Manon Jammes Abbas Tabasi Trung Bach Aideen E. Ryan Thomas Ritter |
| author_sort | Jiemin Wang |
| collection | DOAJ |
| description | Abstract Background Mesenchymal stromal cells (MSCs) possess strong immunomodulatory properties, making them attractive candidates for regenerative medicine and immune-related therapies. Pre-activation, or licensing, of MSCs with cytokines such as interferon-gamma (IFN-γ) and transforming growth factor-beta 1 (TGF-β1) has been shown to enhance their immunosuppressive efficacy. Recent attention has turned to extracellular vesicles (EVs) released by licensed MSCs as a cell-free therapeutic alternative. Methods Small EVs were isolated from MSCs licensed with a combination of IFN-γ and TGF-β1. These EVs were characterized according to standardized criteria. Their immunomodulatory effects were assessed in vitro using two human immune models: a THP-1-derived macrophage polarization system and a peripheral blood mononuclear cell (PBMC) co-culture assay. Pro/anti-inflammatory molecules secretion, T cell proliferation, and regulatory T cell induction were quantified. Dimensionality reduction using t-distributed stochastic neighbor embedding (t-SNE) was applied to multiparametric flow cytometry data for immune profiling. In addition, publicly available transcriptomic datasets (GSE122091 and GSE46019) were analyzed to identify differentially expressed genes (DEGs) in IFN-γ– and TGF-β1–licensed MSCs, providing insight into potential molecular drivers of EV-mediated immunoregulation. Results Licensed EVs significantly inhibited pro-inflammatory THP-1 macrophage activation and promoted an anti-inflammatory phenotype, with reduced secretion of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β), increased IL-10 production, and decreased nitric oxide (NO) levels.. Compared to EVs from non-licensed MSCs, licensed EVs induced a greater proportion of regulatory T cells and exhibited enhanced suppression of allogeneic T cell proliferation. t-SNE analysis revealed a distinct immunoregulatory signature induced by licensed EVs, characterized by the emergence of a non-proliferative lymphocyte subset with elevated co-expression of CD4, CD25, and FOXP3. Transcriptomic analysis further revealed seven overlapping DEGs between IFN-γ– and TGF-β1–licensed MSCs, including both upregulated (GPR68, LIMK2, LIPG) and downregulated (EFNA5, PRKG1, DCLK1, TRIM2) genes, several of which are functionally implicated in EV-mediated immune regulation. Conclusions Small EVs derived from IFN-γ and TGF-β1-licensed MSCs exhibit demonstrate dose-dependent immunomodulatory trends in vitro, with enhanced effects observed at higher concentrations.. These findings suggest their potential utility in modulating both innate and adaptive immune responses, warranting further investigation for their application as a cell-free therapeutic strategy in immune-mediated conditions. Graphic Abstract |
| format | Article |
| id | doaj-art-65a8cf0aa8724e309c7f7d56bfcfaf3a |
| institution | DOAJ |
| issn | 1757-6512 |
| language | English |
| publishDate | 2025-07-01 |
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| spelling | doaj-art-65a8cf0aa8724e309c7f7d56bfcfaf3a2025-08-20T03:04:26ZengBMCStem Cell Research & Therapy1757-65122025-07-0116111910.1186/s13287-025-04476-2In-vitro immunomodulatory efficacy of extracellular vesicles derived from TGF-β1/IFN-γ dual licensed human bone marrow mesenchymal stromal cellsJiemin Wang0Seyedmohammad Moosavizadeh1Manon Jammes2Abbas Tabasi3Trung Bach4Aideen E. Ryan5Thomas Ritter6School of Medicine, Biomedical Sciences, Regenerative Medicine Institute, University of GalwaySchool of Medicine, Biomedical Sciences, Regenerative Medicine Institute, University of GalwaySchool of Medicine, Biomedical Sciences, Regenerative Medicine Institute, University of GalwaySchool of Medicine, Biomedical Sciences, Regenerative Medicine Institute, University of GalwaySchool of Medicine, Biomedical Sciences, Regenerative Medicine Institute, University of GalwaySchool of Medicine, Biomedical Sciences, Regenerative Medicine Institute, University of GalwaySchool of Medicine, Biomedical Sciences, Regenerative Medicine Institute, University of GalwayAbstract Background Mesenchymal stromal cells (MSCs) possess strong immunomodulatory properties, making them attractive candidates for regenerative medicine and immune-related therapies. Pre-activation, or licensing, of MSCs with cytokines such as interferon-gamma (IFN-γ) and transforming growth factor-beta 1 (TGF-β1) has been shown to enhance their immunosuppressive efficacy. Recent attention has turned to extracellular vesicles (EVs) released by licensed MSCs as a cell-free therapeutic alternative. Methods Small EVs were isolated from MSCs licensed with a combination of IFN-γ and TGF-β1. These EVs were characterized according to standardized criteria. Their immunomodulatory effects were assessed in vitro using two human immune models: a THP-1-derived macrophage polarization system and a peripheral blood mononuclear cell (PBMC) co-culture assay. Pro/anti-inflammatory molecules secretion, T cell proliferation, and regulatory T cell induction were quantified. Dimensionality reduction using t-distributed stochastic neighbor embedding (t-SNE) was applied to multiparametric flow cytometry data for immune profiling. In addition, publicly available transcriptomic datasets (GSE122091 and GSE46019) were analyzed to identify differentially expressed genes (DEGs) in IFN-γ– and TGF-β1–licensed MSCs, providing insight into potential molecular drivers of EV-mediated immunoregulation. Results Licensed EVs significantly inhibited pro-inflammatory THP-1 macrophage activation and promoted an anti-inflammatory phenotype, with reduced secretion of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β), increased IL-10 production, and decreased nitric oxide (NO) levels.. Compared to EVs from non-licensed MSCs, licensed EVs induced a greater proportion of regulatory T cells and exhibited enhanced suppression of allogeneic T cell proliferation. t-SNE analysis revealed a distinct immunoregulatory signature induced by licensed EVs, characterized by the emergence of a non-proliferative lymphocyte subset with elevated co-expression of CD4, CD25, and FOXP3. Transcriptomic analysis further revealed seven overlapping DEGs between IFN-γ– and TGF-β1–licensed MSCs, including both upregulated (GPR68, LIMK2, LIPG) and downregulated (EFNA5, PRKG1, DCLK1, TRIM2) genes, several of which are functionally implicated in EV-mediated immune regulation. Conclusions Small EVs derived from IFN-γ and TGF-β1-licensed MSCs exhibit demonstrate dose-dependent immunomodulatory trends in vitro, with enhanced effects observed at higher concentrations.. These findings suggest their potential utility in modulating both innate and adaptive immune responses, warranting further investigation for their application as a cell-free therapeutic strategy in immune-mediated conditions. Graphic Abstracthttps://doi.org/10.1186/s13287-025-04476-2Mesenchymal stromal cells (MSCs)Cytokine licensingExtracellular vesicles (EVs)ImmunomodulationRegulatory T cells (Treg)t-distributed stochastic neighbor embedding (t-SNE) analysis |
| spellingShingle | Jiemin Wang Seyedmohammad Moosavizadeh Manon Jammes Abbas Tabasi Trung Bach Aideen E. Ryan Thomas Ritter In-vitro immunomodulatory efficacy of extracellular vesicles derived from TGF-β1/IFN-γ dual licensed human bone marrow mesenchymal stromal cells Stem Cell Research & Therapy Mesenchymal stromal cells (MSCs) Cytokine licensing Extracellular vesicles (EVs) Immunomodulation Regulatory T cells (Treg) t-distributed stochastic neighbor embedding (t-SNE) analysis |
| title | In-vitro immunomodulatory efficacy of extracellular vesicles derived from TGF-β1/IFN-γ dual licensed human bone marrow mesenchymal stromal cells |
| title_full | In-vitro immunomodulatory efficacy of extracellular vesicles derived from TGF-β1/IFN-γ dual licensed human bone marrow mesenchymal stromal cells |
| title_fullStr | In-vitro immunomodulatory efficacy of extracellular vesicles derived from TGF-β1/IFN-γ dual licensed human bone marrow mesenchymal stromal cells |
| title_full_unstemmed | In-vitro immunomodulatory efficacy of extracellular vesicles derived from TGF-β1/IFN-γ dual licensed human bone marrow mesenchymal stromal cells |
| title_short | In-vitro immunomodulatory efficacy of extracellular vesicles derived from TGF-β1/IFN-γ dual licensed human bone marrow mesenchymal stromal cells |
| title_sort | in vitro immunomodulatory efficacy of extracellular vesicles derived from tgf β1 ifn γ dual licensed human bone marrow mesenchymal stromal cells |
| topic | Mesenchymal stromal cells (MSCs) Cytokine licensing Extracellular vesicles (EVs) Immunomodulation Regulatory T cells (Treg) t-distributed stochastic neighbor embedding (t-SNE) analysis |
| url | https://doi.org/10.1186/s13287-025-04476-2 |
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