{"doi":"10.1111/cei.13537","title":"CD40L-stimulated B cells for<i>ex-vivo</i>expansion of polyspecific non-human primate regulatory T cells for translational studies","abstract":"Summary The therapeutic applications of regulatory T cells (Tregs) include treating autoimmune diseases, graft-versus-host disease and induction of transplantation tolerance. For ex-vivo expanded Tregs to be used in deceased donor transplantation, they must be able to suppress T cell responses to a broad range of human leukocyte antigen (HLA). Here, we present a novel approach for the expansion of polyspecific Tregs in cynomolgus macaques that was adapted from a good manufacturing practice-compliant protocol. Tregs were isolated by fluorescence-activated cell sorting (FACS) and expanded in the presence of a panel of CD40L-stimulated B cells (CD40L-sBc). Prior to Treg culture, CD40L-sBc were expanded in vitro from multiple major histocompatibility complex (MHC)-disparate macaques. Expanded Tregs expressed high levels of forkhead box protein 3 (FoxP3) and Helios, a high percentage of Treg-specific demethylated region (TSDR) demethylation and strong suppression of naïve T cell responses in vitro. In addition, these Tregs produced low levels of inflammatory cytokines and were able to expand post-cryopreservation. Specificity assays confirmed that these Tregs were suppressive upon activation by any antigen-presenting cells (APCs) whose MHC was shared by CD40L-sBc used during expansion, proving that they are polyspecific. We developed an approach for the expansion of highly suppressive cynomolgus macaque polyspecific Tregs through the use of a combination of CD40L-engineered B cells with the potential to be translated to clinical studies. To our knowledge, this is the first report that uses a pool of MHC-mismatched CD40L-sBc to create polyspecific Tregs suitable for use in deceased-donor transplants.","journal":"Clinical & Experimental Immunology","year":2020,"id":107610,"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":6,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9526,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":517238,"name":"Nathaly Llore","orcid":null,"position":1,"is_corresponding":false},{"id":517239,"name":"E. R. Lopes","orcid":null,"position":2,"is_corresponding":false},{"id":517240,"name":"S-B Kofman","orcid":null,"position":3,"is_corresponding":false},{"id":517241,"name":"S-H Ho","orcid":null,"position":4,"is_corresponding":false},{"id":49798,"name":"Jeffrey Stern","orcid":"0000-0001-5588-0357","position":5,"is_corresponding":false},{"id":517242,"name":"Gillotay Pierre","orcid":null,"position":6,"is_corresponding":false},{"id":517243,"name":"Karina Bruestle","orcid":null,"position":7,"is_corresponding":false},{"id":285543,"name":"Qizhi Tang","orcid":"0000-0001-7313-3574","position":8,"is_corresponding":false},{"id":369810,"name":"Megan Sykes","orcid":"0000-0002-4947-4376","position":9,"is_corresponding":false},{"id":49824,"name":"Adam Griesemer","orcid":"0000-0003-4894-2024","position":10,"is_corresponding":false},{"id":402551,"name":"Paula Alonso‐Guallart","orcid":"0000-0002-8801-2457","position":0,"is_corresponding":true}],"reference_count":34,"raw_metadata":null,"created_at":"2026-07-18T23:12:34.898963Z","pmid":"33058141","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":[]}