Author:
Brown Matthew E.,Peters Leeana D.,Hanbali Seif R.,Arnoletti Juan M.,Sachs Lindsey K.,Nguyen Kayla Q.,Carpenter Emma B.,Seay Howard R.,Fuhrman Christopher A.,Posgai Amanda L.,Shapiro Melanie R.,Brusko Todd M.
Abstract
Regulatory T cell (Treg) adoptive cell therapy (ACT) represents an emerging strategy for restoring immune tolerance in autoimmune diseases. Tregs are commonly purified using a CD4+CD25+CD127lo/-gating strategy, which yields a mixed population: 1) cells expressing the transcription factors, FOXP3 and Helios, that canonically define lineage stable thymic Tregs and 2) unstable FOXP3+Helios-Tregs. Our prior work identified the autoimmune disease risk-associated locus and costimulatory molecule, CD226, as being highly expressed not only on effector T cells but also, interferon-γ (IFN-γ) producing peripheral Tregs (pTreg). Thus, we sought to determine whether isolating Tregs with a CD4+CD25+CD226-strategy yields a population with increased purity and suppressive capacity relative to CD4+CD25+CD127lo/-cells. After 14d of culture, expanded CD4+CD25+CD226-cells displayed a decreased proportion of pTregs relative to CD4+CD25+CD127lo/-cells, as measured by FOXP3+Helios-expression and the epigenetic signature at theFOXP3Treg-specific demethylated region (TSDR). Furthermore, CD226-Tregs exhibited decreased production of the effector cytokines, IFN-γ, TNF, and IL-17A, along with increased expression of the immunoregulatory cytokine, TGF-β1. Lastly, CD226-Tregs demonstrated increasedin vitrosuppressive capacity as compared to their CD127lo/-counterparts. These data suggest that the exclusion of CD226-expressing cells during Treg sorting yields a population with increased purity, lineage stability, and suppressive capabilities, which may benefit Treg ACT for the treatment of autoimmune diseases.
Funder
National Institutes of Health
Leona M. and Harry B. Helmsley Charitable Trust
Subject
Immunology,Immunology and Allergy