Primary Human Lymphocytes Transduced with NY-ESO-1 Antigen-Specific TCR Genes Recognize and Kill Diverse Human Tumor Cell Lines

摘要: cDNAs encoding TCR α- and β-chains specific for HLA-A2-restricted cancer-testis Ag NY-ESO-1 were cloned using a 5′RACE method from RNA isolated CTL generated by in vitro stimulation of PBMC with modified NY-ESO-1-specific peptide (p157–165, 9V). Functionality the was confirmed electroporation primary PBL. cDNA these used to construct murine stem cell virus-based retroviral vector, high titer packaging lines generated. Gene transfer efficiency T lymphocytes up 60% obtained without selection precoating vectors onto culture plates. Both CD4+ CD8+ cells could be transduced at same efficiency. High avidity recognition demonstrated coculture target pulsed low levels (<20 pM). TCR-transduced CD4 cells, when cocultured T2 produce IFN-γ, GM-CSF, IL-4, IL-10, suggesting CD8-independent, activation. The efficiently recognize kill HLA-A2- NY-ESO-1-positive melanoma 4-h 51Cr release assay. Finally, nonmelanoma tumor lines. These results strongly support idea that redirection normal specificity gene can have potential applications adoptive immunotherapy.