{"doi":"10.3389/fimmu.2022.873560","title":"Human CD4+CD25+CD226- Tregs Demonstrate Increased Purity, Lineage Stability, and Suppressive Capacity Versus CD4+CD25+CD127lo/- Tregs for Adoptive Cell Therapy","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 + CD127 lo/- 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 + CD127 lo/- cells. After 14d of culture, expanded CD4 + CD25 + CD226 - cells displayed a decreased proportion of pTregs relative to CD4 + CD25 + CD127 lo/- cells, as measured by FOXP3 + Helios - expression and the epigenetic signature at the FOXP3 Treg-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 increased in vitro suppressive capacity as compared to their CD127 lo/- 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.","journal":"Frontiers in Immunology","year":2022,"id":245028,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":25,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9537,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":356514,"name":"Leeana D. Peters","orcid":"0000-0002-7878-0014","position":1,"is_corresponding":false},{"id":880164,"name":"Seif R. Hanbali","orcid":null,"position":2,"is_corresponding":false},{"id":879423,"name":"Juan M. Arnoletti","orcid":"0000-0002-1313-8725","position":3,"is_corresponding":false},{"id":880165,"name":"Lindsey K. Sachs","orcid":null,"position":4,"is_corresponding":false},{"id":880166,"name":"Kayla Q. Nguyen","orcid":null,"position":5,"is_corresponding":false},{"id":880167,"name":"Emma B. Carpenter","orcid":null,"position":6,"is_corresponding":false},{"id":356515,"name":"Howard R. Seay","orcid":"0000-0001-7002-0187","position":7,"is_corresponding":false},{"id":59673,"name":"Christopher A. Fuhrman","orcid":"0000-0002-5364-6935","position":8,"is_corresponding":false},{"id":251267,"name":"Amanda L. Posgai","orcid":"0000-0002-9491-0958","position":9,"is_corresponding":false},{"id":479037,"name":"Melanie R. Shapiro","orcid":"0000-0003-2090-0877","position":10,"is_corresponding":false},{"id":356521,"name":"Todd M. Brusko","orcid":"0000-0003-2878-9296","position":11,"is_corresponding":false},{"id":729988,"name":"Matthew E. Brown","orcid":"0000-0001-8230-5243","position":0,"is_corresponding":true}],"reference_count":79,"raw_metadata":null,"created_at":"2026-07-19T00:23:34.974895Z","pmid":"35693814","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}