title: rAAV/Her-2/neu loading of dendritic cells for a potent cellular-mediated MHC class I restricted immune response against ovarian cancer.
abstract: Recent studies demonstrate that recombinant adeno-associated virus (rAAV)-based antigen loading of dendritic cells (DCs) generates, in vitro, significant and rapid cytotoxic T-lymphocyte (CTL) responses against viral antigens. We used the rAAV system to induce specific CTLs against tumor antigens for the development of ovarian cancer (OC) gene therapy. As an extension of the versatility of the rAAV system, we incorporated a self-antigen, Her-2/neu, which is expressed in many cancers, including breast and ovarian. We analyzed two different vectors containing a short (157-612) and long domain (1-1197). Our rAAV vector induced strong stimulation of CTLs directed against the self tumor antigen, Her-2/neu. We then investigated the efficiency of the CTLs in killing Her-2/neu-targeted cells. A significant MHC class I-restricted, anti-Her-2/neu-specific CTL killing was demonstrated against Her-2/neu-positive OC cells after one in vitro stimulation. In summary, single peripheral blood mononuclear cell (PBMC) stimulation with rAAV/157-612- or rAAV/1-1197-pulsed DCs induces strong antigen-specific CTL generation. The CTLs were capable of lysing low doses of peptides pulsed into target cells or OC Her-2/neu(+) tumors. These data suggest that AAV-based antigen loading of DCs is highly effective for generating human CTL responses against OC antigens.
authors.affiliation: Division of Hematology and Oncology, Texas Tech University Health Sciences Center and the Southwest Cancer Treatment and Research Center, Lubbock, Texas 79430, USA.
authors.forename: Yuefei
authors.initials: Y
authors.lastname: Yu
authors.forename: Petra
authors.initials: P
authors.lastname: Pilgrim
authors.forename: Wei
authors.initials: W
authors.lastname: Zhou
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Gagliano
authors.forename: E Eldo
authors.initials: EE
authors.lastname: Frezza
authors.forename: Marjorie
authors.initials: M
authors.lastname: Jenkins
authors.forename: Jon A
authors.initials: JA
authors.lastname: Weidanz
authors.forename: Joseph
authors.initials: J
authors.lastname: Lustgarten
authors.forename: Martin
authors.initials: M
authors.lastname: Cannon
authors.forename: Klaus
authors.initials: K
authors.lastname: Bumm
authors.forename: Everardo
authors.initials: E
authors.lastname: Cobos
authors.forename: W Martin
authors.initials: WM
authors.lastname: Kast
authors.forename: Maurizio
authors.initials: M
authors.lastname: Chiriva-Internati
mesh_terms: D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic; D003713:Dendritic Cells / Q000276:immunology* / Q000821:virology; D000229:Dependovirus / Q000235:genetics* / Q000276:immunology; D005260:Female; D015870:Gene Expression; D005805:Genes, MHC Class I; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology; D006801:Humans; D007111:Immunity, Cellular; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000821:virology; D010051:Ovarian Neoplasms / Q000235:genetics / Q000276:immunology* / Q000821:virology; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000821:virology
chemical_list: D000951:Antigens, Neoplasm; D018719:Erb-b2 Receptor Tyrosine Kinases