{"doi":"10.1128/mbio.03413-23","title":"A novel lentiviral vector-based approach to generate chimeric antigen receptor T cells targeting <i>Aspergillus fumigatus</i>","abstract":"ABSTRACT Invasive aspergillosis (IA) is a common and deadly mold infection in immunocompromised patients. As morbidity and mortality of IA are primarily driven by poor immune defense, adjunct immunotherapies, such as chimeric antigen receptor (CAR) T cells, are direly needed. Here, we propose a novel approach to generate Aspergillus fumigatus (AF)-CAR T cells using the single-chain variable fragment domain of monoclonal antibody AF-269-5 and a lentiviral vector system. These cells successfully targeted mature hyphal filaments of representative clinical and reference AF isolates and elicited a potent release of cytotoxic effectors and type 1 T cell cytokines. Furthermore, AF-CAR T cells generated from peripheral blood mononuclear cells of four healthy human donors and expanded with either of three cytokine stimulation regimens (IL-2, IL-2 + IL-21, or IL-7 + IL-15) significantly suppressed mycelial growth of AF-293 after 18 hours of co-culture and synergized with the immunomodulatory antifungal agent caspofungin to control hyphal growth for 36 hours. Moreover, cyclophosphamide-immunosuppressed NSG mice with invasive pulmonary aspergillosis that received two doses of 5 million AF-CAR T cells (6 and 48 hours after AF infection) showed significantly reduced morbidity on day 4 post-infection ( P &lt; 0.001) and significantly improved 7-day survival ( P = 0.049) compared with mice receiving non-targeting control T cells, even without concomitant antifungal chemotherapy. In conclusion, we developed a novel lentiviral strategy to obtain AF-CAR T cells with high targeting efficacy, yielding significant anti-AF activity in vitro and short-term protection in vivo . Our approach could serve as an important steppingstone for future clinical translation of antifungal CAR T-cell therapy after further refinement and thorough preclinical evaluation. IMPORTANCE Invasive aspergillosis (IA) remains a formidable cause of morbidity and mortality in patients with hematologic malignancies and those undergoing hematopoietic stem cell transplantation. Despite the introduction of several new Aspergillus -active antifungals over the last 30 years, the persisting high mortality of IA in the setting of continuous and profound immunosuppression is a painful reminder of the major unmet need of effective antifungal immune enhancement therapies. The success of chimeric antigen receptor (CAR) T-cell therapy in cancer medicine has inspired researchers to translate this approach to opportunistic infections, including IA. Aiming to refine anti- Aspergillus CAR T-cell therapy and improve its feasibility for future clinical translation, we herein developed and validated a novel antibody-based CAR construct and lentiviral transduction method to accelerate the production of CAR T cells with high targeting efficacy against Aspergillus fumigatus . Our unique approach could provide a promising platform for future clinical translation of CAR T-cell-based antifungal immunotherapy.","journal":"mBio","year":2024,"id":425891,"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":18,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9497,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":301203,"name":"Sebastian Wurster","orcid":"0000-0002-7784-7256","position":1,"is_corresponding":false},{"id":897823,"name":"Karishma Bavisi","orcid":null,"position":2,"is_corresponding":false},{"id":1224162,"name":"Thiago Aparecido da Silva","orcid":"0000-0001-5017-6539","position":3,"is_corresponding":false},{"id":1224726,"name":"Paul Hauser","orcid":null,"position":4,"is_corresponding":false},{"id":1224727,"name":"Jordan Kinnitt","orcid":null,"position":5,"is_corresponding":false},{"id":425409,"name":"Nathaniel D. Albert","orcid":null,"position":6,"is_corresponding":false},{"id":235090,"name":"Uddalak Bharadwaj","orcid":"0000-0002-8676-7258","position":7,"is_corresponding":false},{"id":11096,"name":"Sattva S. Neelapu","orcid":"0000-0003-1045-4914","position":8,"is_corresponding":false},{"id":95683,"name":"Dimitrios P. Kontoyiannis","orcid":"0000-0002-8051-2940","position":9,"is_corresponding":false},{"id":1182805,"name":"Pappanaicken R. Kumaresan","orcid":null,"position":0,"is_corresponding":true}],"reference_count":39,"raw_metadata":null,"created_at":"2026-07-19T01:58:32.160975Z","pmid":"38415653","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":[]}