Extracellular Vesicles isolated from Mesenchymal Stromal Cells Modulate CD4+ T Lymphocytes Toward a Regulatory Profile.

Flavia Franco Da Cunha,Regiane Aparecida Cavinato,Mario Costa Cruz,Vinicius Andrade-Oliveira,Danilo Candido De Almeida,Niels Olsen Saraiva Câmara,Alvaro Pacheco-Silva,Cristiane Naffah De Souza Breda,Tamiris Borges Da Silva,Marcos Antonio Cenedeze,Ana Claudia Torrecilhas,Meire Ioshie Hiyane,Eliana L Faquim-Mauro

doi:10.3390/cells9041059

Abstract

Mesenchymal stromal cells (MSCs) can generate immunological tolerance due to their regulatory activity in many immune cells. Extracellular vesicles (EVs) release is a pivotal mechanism by which MSCs exert their actions. In this study, we evaluate whether mesenchymal stromal cell extracellular vesicles (MSC-EVs) can modulate T cell response. MSCs were expanded and EVs were obtained by differential ultracentrifugation of the supernatant. The incorporation of MSC-EVs by T cells was detected by confocal microscopy. Expression of surface markers was detected by flow cytometry or CytoFLEX and cytokines were detected by RT-PCR, FACS and confocal microscopy and a miRNA PCR array was performed. We demonstrated that MSC-EVs were incorporated by lymphocytes in vitro and decreased T cell proliferation and Th1 differentiation. Interestingly, in Th1 polarization, MSC-EVs increased Foxp3 expression and generated a subpopulation of IFN-γ+/Foxp3+T cells with suppressive capacity. A differential expression profile of miRNAs in MSC-EVs-treated Th1 cells was seen, and also a modulation of one of their target genes, TGFbR2. MSC-EVs altered the metabolism of Th1-differentiated T cells, suggesting the involvement of the TGF-β pathway in this metabolic modulation. The addition of MSC-EVs in vivo, in an OVA immunization model, generated cells Foxp3+. Thus, our findings suggest that MSC-EVs are able to specifically modulate activated T cells at an alternative regulatory profile by miRNAs and metabolism shifting.

Highlights

Mesenchymal stromal cells (MSCs) are adherent cells, capable of proliferating and differentiating in mature cells of mesenchymal lines [1] and expressing CD73, CD90, CD105 [2]
We demonstrate the ability of MSC-extracellular vesicles (EVs) to regulate Th1 cells potentially via modulation of miRNA profile associated TGF-β pathway and metabolism shifting
We showed a reduction in glycolytic and mitochondrial metabolism in the cells differentiated in the presence of mesenchymal stromal cell extracellular vesicles (MSC-EVs)

Summary

Introduction

Mesenchymal stromal cells (MSCs) are adherent cells, capable of proliferating and differentiating in mature cells of mesenchymal lines [1] and expressing CD73, CD90, CD105 [2]. Studies that pursue to identify the mechanisms by which MSCs exert their regulation suggest that the paracrine effect is more important than cell-cell contact, being the main mediator of this action [3]. In this context, the release of soluble factors with immunomodulatory properties, such as HGF [11], TGF-β [7], IL-10 [12], prostaglandin-E2 (PGE2 ) [13], indoleamine-2,3-dioxygenase (IDO) [14], has been identified as responsible for the effects of MSCs in several studies. The release of extracellular vesicles (EVs) by these cells has been demonstrated as an alternative mechanism by which MSCs perform their biologic effects [15]

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Results

Conclusion