{"doi":"10.1101/2022.11.14.516164","title":"Phenotypic, molecular and functional characterisation of human <i>in vitro</i> -generated IL-17A+ CD8+ T-cells","abstract":"Abstract IL-17A+ CD8+ T-cells, often referred to as Tc17 cells, have been identified at sites of inflammation in several immune-mediated inflammatory diseases including psoriasis and spondyloarthritis. Whilst much of our understanding of IL-17A+ CD8+ T-cells has been discerned from murine studies, human IL-17A+ CD8+ T-cells remain less-well characterised. We optimised an in vitro polarisation protocol to expand human IL-17A+ CD8+ T-cells from PBMC or bulk CD8+ T-cell populations for phenotypic and functional assessment. We show that T-cell activation in the presence of IL-1β and IL-23 significantly increased the frequencies of IL-17A+ CD8+ T-cells, which was not further enhanced by the addition of IL-6, IL-2 or anti-IFNγ mAb. In vitro-generated IL-17A+ CD8+ T-cells from healthy donors displayed a distinct type-17 profile compared with IL-17A- CD8+ T-cells, as defined by transcriptional signature ( IL17A, IL17F, RORC, RORA, MAF, IL23R, CCR6, CXCR6 ); high surface expression of CCR6 and CD161; and polyfunctional production of IL-17A, IL-17F, IL-22, IFNγ, TNFα and GM-CSF. A significant proportion of in vitro -induced IL-17A+ CD8+ T-cells expressed TCRVα7.2 and bound MR1 tetramers, indicative of a MAIT CD8+ T-cell population. Using an IL-17A secretion assay, we demonstrate that the in vitro -generated IL-17A+ CD8+ T-cells were biologically functional and induced pro-inflammatory IL-6 and IL-8 production by synovial fibroblasts from patients with psoriatic arthritis. Collectively, we report an in vitro culture system to expand IL-17A+ CD8+ T-cells and further characterise their phenotype, transcriptional regulation and functional relevance to human health and disease.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2022,"id":304830,"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":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9597,"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":998241,"name":"Ushani Srenathan","orcid":"0000-0002-9877-1991","position":1,"is_corresponding":false},{"id":998242,"name":"Lucy E. Durham","orcid":"0000-0002-6726-9245","position":2,"is_corresponding":false},{"id":998822,"name":"Sylvine Lalnunhlimi","orcid":null,"position":3,"is_corresponding":false},{"id":270788,"name":"Kathryn J. A. Steel","orcid":"0000-0002-4446-0506","position":4,"is_corresponding":false},{"id":28664,"name":"Anca I. Catrina","orcid":"0000-0002-1292-1152","position":5,"is_corresponding":false},{"id":998243,"name":"Bruce Kirkham","orcid":"0000-0002-5110-3320","position":6,"is_corresponding":false},{"id":998244,"name":"Leonie S. Taams","orcid":"0000-0002-9337-7194","position":7,"is_corresponding":false},{"id":998821,"name":"Elizabeth H. Gray","orcid":null,"position":0,"is_corresponding":true}],"reference_count":62,"raw_metadata":null,"created_at":"2026-07-19T00:32:37.185846Z","pmid":null,"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":[]}