CD73-bearing Treg-derived extracellular vesicles modulate in vitro and in vivo immune response and prevent alveolar bone loss during periodontitis
Objectives: The CD73 ectoenzyme promotes immune regulation through the conversion of adenosine-mono phosphate (AMP) into adenosine, which after interaction with its receptors (A2aR and A2bR expressed on immune cells) upholds anti-inflammatory responses. T regulatory cells (Treg)-derived extracellular vesicles (TEVs) have been shown to be loaded with CD73, contributing to their immunomodulatory capacity in a cell-free manner. Periodontitis, a chronic inflammatory disease, develops a deregulated host immune response, where the formation of a inflammatory infiltrate and accumulation of bone pro-resorptive factors result in tooth-supporting tissue breakdown and alveolar bone loss. This study aimed to evaluate the effect of CD73-bearing TEVs on T-cell proliferation, activation, and phenotype in vitro and on periodontitis-induced immune response and alveolar bone resorption. Methods: Magnetically isolated murine CD4+ T cells were differentiated into Tregs and characterized by flow cytometry (FC). From their recovered supernatant, TEVs were isolated by differential centrifugation then, quantified and characterized by Nanoparticle-Tracking-Analysis and Western Blot. The TEV immunosuppressive and modulatory capacities were evaluated in vitro on CD4+ and CD8+ T cells studying their effect on proliferation, activation, and phenotype. Besides, using a ligature-induced periodontitis mice model, the TEVs effect on periodontal immune response was assessed quantifying leukocyte infiltration by FC and the extent of alveolar bone loss by morphometric analysis. Results: Tregs showed high expression of lineage markers Foxp3, CD25, and CD73. Isolated TEVs expressed CD73, displayed canonical EVs characteristics, downregulated the CD4+ and upregulated the CD8+ T cell proliferation and activation. During periodontitis, TEVs increased the frequency of CD73+ leukocytes, promoted anti-inflammatory response, and prevented alveolar bone loss. Conclusions: CD73-bearing TEVs modulate CD4+ and CD8+ T cell response, impacts on local immune response and prevent peridontitis-induced alveolar bone loss.
Rojas Pérez, Carolina
( Universidad de Chile
, Santiago
, Región Metropolitana
, Chile
; Centro de Investigación e Innovación Biomédica (CIIB) Universidad de los Andes
, Santiago
, Region Metropolitana
, Chile
)
García, Michelle
( Universidad de Chile
, Santiago
, Región Metropolitana
, Chile
)
Gonzalez-osuna, Luis
( Universidad de Chile
, Santiago
, Región Metropolitana
, Chile
)
Galvez-jirón, Felipe
( Centro de Investigación e Innovación Biomédica (CIIB) Universidad de los Andes
, Santiago
, Region Metropolitana
, Chile
)
Cafferata, Emilio
( Universidad de Chile
, Santiago
, Región Metropolitana
, Chile
)
Pino-lagos, Karina
( Centro de Investigación e Innovación Biomédica (CIIB) Universidad de los Andes
, Santiago
, Region Metropolitana
, Chile
)
Vernal Astudillo, Rolando
( Universidad de Chile
, Santiago
, Región Metropolitana
, Chile
)
NONE
Proyecto FONDECYT Regular 1181780 y 1210654. Beca ANID 21180841