{"doi":"10.1136/jitc-2023-007803","title":"CD8<sup>+</sup> chimeric antigen receptor T cells manufactured in absence of CD4<sup>+</sup> cells exhibit hypofunctional phenotype","abstract":"Background Cell culture conditions during manufacturing can impact the clinical efficacy of chimeric antigen receptor (CAR) T cell products. Production methods have not been standardized because the optimal approach remains unknown. Separate CD4 + and CD8 + cultures offer a potential advantage but complicate manufacturing and may affect cell expansion and function. In a phase 1/2 clinical trial, we observed poor expansion of separate CD8 + cell cultures and hypothesized that coculture of CD4 + cells and CD8 + cells at a defined ratio at culture initiation would enhance CD8 + cell expansion and simplify manufacturing. Methods We generated CAR T cells either as separate CD4 + and CD8 + cells, or as combined cultures mixed in defined CD4:CD8 ratios at culture initiation. We assessed CAR T cell expansion, phenotype, function, gene expression, and in vivo activity of CAR T cells and compared these between separately expanded or mixed CAR T cell cultures. Results We found that the coculture of CD8 + CAR T cells with CD4 + cells markedly improves CD8 + cell expansion, and further discovered that CD8 + cells cultured in isolation exhibit a hypofunctional phenotype and transcriptional signature compared with those in mixed cultures with CD4 + cells. Cocultured CAR T cells also confer superior antitumor activity in vivo compared with separately expanded cells. The positive impact of CD4 + cells on CD8 + cells was mediated through both cytokines and direct cell contact, including CD40L-CD40 and CD70-CD27 interactions. Conclusions Our data indicate that CD4 + cell help during cell culture maintains robust CD8 + CAR T cell function, with implications for clinical cell manufacturing.","journal":"Journal for ImmunoTherapy of Cancer","year":2023,"id":323801,"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":36,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9601,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":833714,"name":"Dong Hoon Lee","orcid":"0000-0003-1521-133X","position":1,"is_corresponding":false},{"id":286050,"name":"Wei Sun","orcid":"0000-0002-6350-1107","position":2,"is_corresponding":false},{"id":1039667,"name":"Francisco Cervantes-Contreras","orcid":null,"position":3,"is_corresponding":false},{"id":295731,"name":"Ryan Basom","orcid":"0000-0001-6137-1807","position":4,"is_corresponding":false},{"id":21506,"name":"Feinan Wu","orcid":"0000-0002-4962-1874","position":5,"is_corresponding":false},{"id":1039157,"name":"Si Liu","orcid":"0000-0003-1289-7241","position":6,"is_corresponding":false},{"id":1039158,"name":"Richa Hirendra","orcid":"0000-0003-0102-2773","position":7,"is_corresponding":false},{"id":348264,"name":"Hamid Reza Mirzaei","orcid":"0000-0002-3303-2565","position":8,"is_corresponding":false},{"id":1039159,"name":"Shyril O’Steen","orcid":"0000-0002-7444-6270","position":9,"is_corresponding":false},{"id":522226,"name":"Damian J. Green","orcid":"0000-0003-0477-1446","position":10,"is_corresponding":false},{"id":233391,"name":"Mazyar Shadman","orcid":"0000-0002-3365-6562","position":11,"is_corresponding":false},{"id":619111,"name":"Brian G. Till","orcid":"0000-0002-4718-8288","position":12,"is_corresponding":false},{"id":1039156,"name":"Sang Yun Lee","orcid":"0000-0002-3071-2661","position":0,"is_corresponding":true}],"reference_count":51,"raw_metadata":null,"created_at":"2026-07-19T01:08:06.496579Z","pmid":"38251688","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":[]}