title: Retargeting NK92 cells using an HLA-A2-restricted, EBNA3C-specific chimeric antigen receptor.
abstract: Advances in adoptive cell immunotherapy have led to several promising options for cancer patients. Single-chain variable fragments (scFvs) were isolated from a human phage display library by panning on recombinant human leukocyte antigen (HLA)-A2-peptide complexes. A scFv (EBNA Clone 315) specific for HLA-A2 carrying a 10 amino acid peptide (LLDFVRFMGV) derived from the Epstein-Barr virus latent protein EBNA3C was fully characterized. EBNA Clone 315 displayed exquisite specificity toward its targeted T-cell epitope (TCE) and did not cross-react with the free peptide, HLA-A2 complexes, which carried irrelevant peptides, or HLA-A2(-) cells. Furthermore, after engineering into a scFv-Fc fusion protein, we were able to determine its affinity, detection sensitivity, and ability to induce antibody-dependent cellular cytotoxicity (ADCC). As a proof-of-principle, a chimeric antigen receptor (CAR) version of EBNA Clone 315 was used to reprogram NK92MI cells. CAR-expressing NK92MI cells showed highly specific and potent cytotoxicity toward the targeted TCE, with detection sensitivity of approximately 25 molecules and cytolytic capacity threefold greater than scFv-Fc-mediated ADCC. For the first time, we show the successful reprogramming of non-T cells toward a specific TCE using a CAR.
authors: Department of Pediatrics, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
authors: D V
authors: DV
authors: Tassev
authors: M
authors: M
authors: Cheng
authors: N-K V
authors: NK
authors: Cheung
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000276:immunology*; D016466:CHO Cells; D002460:Cell Line; D046528:Chimerism; D006224:Cricetinae; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D019309:Epstein-Barr Virus Nuclear Antigens; D015789:HLA-A2 Antigen / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology; D007694:Killer Cells, Natural / Q000276:immunology*; D009842:Oligopeptides / Q000276:immunology* / Q000378:metabolism; D011946:Receptors, Antigen / Q000235:genetics / Q000276:immunology*
chemical_list: D000956:Antigens, Viral; C498774:EBNA-3C, epstein-barr virus; D018984:Epitopes, T-Lymphocyte; D019309:Epstein-Barr Virus Nuclear Antigens; D015789:HLA-A2 Antigen; D007128:Immunoglobulin Fragments; D009842:Oligopeptides; D011946:Receptors, Antigen