title: Lentivirus vector-mediated expression of tumor-associated epitopes by human antigen presenting cells.
abstract: Directing the human immune system to recognize and eliminate tumor cells is the ultimate goal of cancer immunotherapy. Vaccinating patients with autologous antigen presenting cells (APC) expressing tumor-associated antigens (TAA) represents a promising approach for activating tumor-reactive T cells in vivo. In addition, APC expressing TAA provide a means of generating tumor-specific T cells in vitro, for therapeutic and diagnostic applications. Lentiviral vectors are attractive vehicles for introducing TAA-encoding genes into APC. In this study, lentiviral vectors expressing the reporter gene GFP or the melanoma-associated antigen tyrosinase were used to transduce three different kinds of human APC: monocyte-derived dendritic cells (DC), CD40L-activated B lymphocytes, and Epstein Barr virus (EBV)-transformed B lymphocytes. Using optimized transduction conditions for each cell type, tyrosinase was expressed at levels sufficient to stimulate antigen-specific major histocompatibility complex (MHC) class I-restricted T cells from melanoma patients. While transduced EBV-B cells demonstrated the highest level of transgene expression, optimal T-cell recognition was achieved with transduced DC. Substituting the CAG promoter for PGK in lentiviral constructs enhanced transgene expression in DC and EBV-B cells, amplifying T cell recognition. Lentiviruses inducing sustained transgene expression with relatively low cellular toxicity and background viral gene expression may be ideal vectors for immunotherapeutic applications.
authors.affiliation: Surgery Branch, National Cancer Institute, Center for Cancer Research, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Gregory
authors.initials: G
authors.lastname: Lizée
authors.forename: Monica I
authors.initials: MI
authors.lastname: Gonzales
authors.forename: Suzanne L
authors.initials: SL
authors.lastname: Topalian
mesh_terms: D000938:Antigen-Presenting Cells / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000276:immunology* / Q000378:metabolism; D001402:B-Lymphocytes / Q000276:immunology; D023201:CD40 Ligand / Q000378:metabolism; D002472:Cell Transformation, Viral; D003713:Dendritic Cells / Q000276:immunology / Q000502:physiology* / Q000821:virology; D000939:Epitopes; D018014:Gene Transfer Techniques; D005805:Genes, MHC Class I; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008164:Luminescent Proteins / Q000378:metabolism; D008545:Melanoma / Q000235:genetics / Q000473:pathology; D058950:Melanoma-Specific Antigens; D009000:Monocytes / Q000166:cytology / Q000821:virology; D014442:Monophenol Monooxygenase / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology; D014161:Transduction, Genetic; D019076:Transgenes / Q000502:physiology
chemical_list: D000951:Antigens, Neoplasm; D000939:Epitopes; D008164:Luminescent Proteins; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins; D023201:CD40 Ligand; D049452:Green Fluorescent Proteins; D014442:Monophenol Monooxygenase