Human thymic Treg-derived extracellular vesicles display a defined proteomic and miRNA signature with donor-variable suppressive activity.
8 affiliations
- 1Laboratory of Immune-Regulation (LIR), Gregorio Marañón Health Research Institute (IiSGM), Hospital General Gregorio Marañon, C/Dr. Esquerdo, 46, 28007, Madrid, Spain.
- 2HepatoGastro Lab, Instituto de Investigación Sanitaria Gregorio Marañón (IiSGM), 28007, Madrid, Spain.
- 3Group of Advanced Immuno-Regulation (GIRA), Gregorio Marañón Health Research Institute (IiSGM), Hospital General Gregorio Marañon, C/Dr. Esquerdo, 46, 28007, Madrid, Spain.
- 4Paediatric Cardiac Surgery Unit of Gregorio, Marañón University Hospital (HGUGM), C/Dr. Esquerdo, 46, 28007, Madrid, Spain.
- 5Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBEREHD), Madrid, Spain.
- 6Unidad de Investigación Materno Infantil Fundación Familia Alonso (UDIMIFFA), Gregorio Marañón Health Research Institute (IiSGM), Madrid, Spain.
- 7Laboratory of Immune-Regulation (LIR), Gregorio Marañón Health Research Institute (IiSGM), Hospital General Gregorio Marañon, C/Dr. Esquerdo, 46, 28007, Madrid, Spain. [email protected].
- 8Group of Advanced Immuno-Regulation (GIRA), Gregorio Marañón Health Research Institute (IiSGM), Hospital General Gregorio Marañon, C/Dr. Esquerdo, 46, 28007, Madrid, Spain. [email protected].
Abstract
Background
Extracellular vesicles (EVs) are key mediators of intercellular communication that transfer functional molecules reflective of their parent cells and are increasingly explored as cell-free therapeutic agents. Immunoregulatory cell types, including regulatory T cells (Tregs), produce EVs with potential anti-inflammatory properties. While thymus-derived Tregs (thyTregs) exhibit high stability and potent suppressive activity, their capacity to generate functional EVs remains unknown.Methods
EVs were isolated by size exclusion chromatography or by immunocapture from ex vivo-expanded human thyTregs and characterized by electron microscopy, nanoparticle tracking analysis, and Western blotting. Proteomic and miRNA transcriptomic analyses were performed to define EV cargo. Functional activity was assessed by co-culturing EVs with activated allogeneic peripheral blood lymphocytes (PBLs) and evaluating proliferation, activation markers, and cytokine production.Results
Tetraspanin-positive ThyTreg-derived EVs exhibited typical vesicular morphology, a mean size of ~ 200 nm, and expression of canonical EV markers (CD9, TSG101). Proteomic profiling identified 314 proteins enriched in immune-related pathways, including CD25, CTLA-4, TGF-β1, ICOS, and granzyme B, reflecting the parental Treg phenotype. Transcriptomic analysis revealed a distinct miRNA signature dominated by let-7 family members and miR-148a-3p, with predicted targets involved in TGF-β/BMP and MAPK signaling. Functionally, thyTreg-EVs suppressed CD4⁺ and CD8⁺ T cell proliferation in a dose-dependent but heterogeneous manner and, in some donors, induced responder cell death. No significant changes in activation marker expression or cytokine production were observed.Conclusions
Human thyTregs release EVs with immunosuppressive properties and a defined molecular cargo. These findings identify thyTreg-derived EVs as a novel, cell-free platform with potential for immunomodulatory therapies.
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BibTeX
@article{fernandezcastillo2026human,
title = {{Human thymic Treg-derived extracellular vesicles display a defined proteomic and miRNA signature with donor-variable suppressive activity.}},
author = {Marta Fernández-Castillo and Elena Blázquez-López and Esther Bernaldo-de-Quiros and Marta Martínez-Bonet and Jorge Gallego-Valle and Ana Pita and Javier Vaquero and Perceval Vellosillo and Rafael Correa-Rocha and Marjorie Pion},
journal = {Molecular medicine (Cambridge, Mass.)},
year = {2026},
month = aug,
doi = {10.1186/s10020-026-01576-4},
url = {https://europepmc.org/article/MED/42972221},
note = {CC BY},
}
RIS
TY - JOUR
TI - Human thymic Treg-derived extracellular vesicles display a defined proteomic and miRNA signature with donor-variable suppressive activity.
AU - Marta Fernández-Castillo
AU - Elena Blázquez-López
AU - Esther Bernaldo-de-Quiros
AU - Marta Martínez-Bonet
AU - Jorge Gallego-Valle
AU - Ana Pita
AU - Javier Vaquero
AU - Perceval Vellosillo
AU - Rafael Correa-Rocha
AU - Marjorie Pion
PY - 2026
DA - 2026/08/06
JO - Molecular medicine (Cambridge, Mass.)
DO - 10.1186/s10020-026-01576-4
UR - https://europepmc.org/article/MED/42972221
AB - Background Extracellular vesicles (EVs) are key mediators of intercellular communication that transfer functional molecules reflective of their parent cells and are increasingly explored as cell-free therapeutic agents. Immunoregulatory cell types, including regulatory T cells (Tregs), produce EVs with potential anti-inflammatory properties. While thymus-derived Tregs (thyTregs) exhibit high stability and potent suppressive activity, their capacity to generate functional EVs remains unknown.Methods EVs were isolated by size exclusion chromatography or by immunocapture from ex vivo-expanded human thyTregs and characterized by electron microscopy, nanoparticle tracking analysis, and Western blotting. Proteomic and miRNA transcriptomic analyses were performed to define EV cargo. Functional activity was assessed by co-culturing EVs with activated allogeneic peripheral blood lymphocytes (PBLs) and evaluating proliferation, activation markers, and cytokine production.Results Tetraspanin-positive ThyTreg-derived EVs exhibited typical vesicular morphology, a mean size of ~ 200 nm, and expression of canonical EV markers (CD9, TSG101). Proteomic profiling identified 314 proteins enriched in immune-related pathways, including CD25, CTLA-4, TGF-β1, ICOS, and granzyme B, reflecting the parental Treg phenotype. Transcriptomic analysis revealed a distinct miRNA signature dominated by let-7 family members and miR-148a-3p, with predicted targets involved in TGF-β/BMP and MAPK signaling. Functionally, thyTreg-EVs suppressed CD4⁺ and CD8⁺ T cell proliferation in a dose-dependent but heterogeneous manner and, in some donors, induced responder cell death. No significant changes in activation marker expression or cytokine production were observed.Conclusions Human thyTregs release EVs with immunosuppressive properties and a defined molecular cargo. These findings identify thyTreg-derived EVs as a novel, cell-free platform with potential for immunomodulatory therapies.
SN - 1076-1551
SN - 1528-3658
ER -