title: Dendritic cells efficiently acquire and present antigen derived from lung cancer cells and induce antigen-specific T-cell responses.
abstract: Active immunotherapy of cancer requires the availability of a source of tumor antigens. To date, no such antigen associated with lung cancer has been identified. We have therefore investigated the ability of dendritic cells (DC) to capture whole irradiated human lung tumor cells and to present a defined surrogate antigen derived from the ingested tumor cells. We also describe an in vitro system using a modified human adenocarcinoma cell line (A549-M1) that expresses the well-characterized, immunogenic influenza M1 matrix protein as a surrogate tumor antigen. Peripheral blood monocyte-derived DC, when co-cultured with sub-lethally irradiated A549 cells or primary lung tumor cells derived from surgical resection of non-small cell carcinoma (NSCLC), efficiently ingested the tumor cells as determined by flow cytometry analysis and confocal microscopic examination. More importantly, DC loaded with irradiated A549-M1 cells efficiently processed and presented tumor cell-derived M1 antigen to T cells and elicited antigen-specific immune responses that included IFNgamma release from an M1-specific T-cell line, expansion of M1 peptide-specific Vbeta17+ and CD8+ peripheral T cells and generation of M1-specific cytotoxic T lymphocytes (CTL). We also compared DC loaded with irradiated tumor cells to those loaded with tumor cell lysate or killed tumor cells and found that irradiated lung tumor cells as a source of tumor antigen for DC loading is superior to tumor cell lysate or killed tumor cells in efficient induction of antigen-specific T-cell responses. Our results demonstrate the feasibility of using lung tumor cell-loaded DC to induce immune responses against lung cancer-associated antigens and support ongoing efforts to develop a DC-based lung cancer vaccine.
authors.affiliation: Vaccine Research and Development Division, Northwest Biotherapeutics Inc., 21720 23rd Drive SE, Suite 100, Bothell, WA 98021, USA. yaling@nwbio.com
authors.forename: Yaling
authors.initials: Y
authors.lastname: Zhou
authors.forename: Julie A
authors.initials: JA
authors.lastname: McEarchern
authors.forename: Edward
authors.initials: E
authors.lastname: Howard
authors.forename: Gary
authors.initials: G
authors.lastname: Pestano
authors.forename: Michael L
authors.initials: ML
authors.lastname: Salgaller
authors.forename: Marnix L
authors.initials: ML
authors.lastname: Bosch
mesh_terms: D000230:Adenocarcinoma / Q000276:immunology; D000256:Adenoviridae / Q000235:genetics; D017951:Antigen Presentation / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000276:immunology*; D015153:Blotting, Western; D002289:Carcinoma, Non-Small-Cell Lung / Q000276:immunology; D018920:Coculture Techniques; D016207:Cytokines / Q000096:biosynthesis; D003601:Cytotoxicity Tests, Immunologic; D003713:Dendritic Cells / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D006801:Humans; D009980:Influenza A virus / Q000276:immunology; D008175:Lung Neoplasms / Q000276:immunology*; D008213:Lymphocyte Activation; D018613:Microscopy, Confocal; D013601:T-Lymphocytes / Q000276:immunology*; D014407:Tumor Cells, Cultured / Q000528:radiation effects; D014763:Viral Matrix Proteins / Q000276:immunology
chemical_list: D000951:Antigens, Neoplasm; D016207:Cytokines; C031540:M-protein, influenza virus; C488936:M1 protein, Influenza A virus; D014763:Viral Matrix Proteins