title: Efficient lysis of epithelial ovarian cancer cells by MAGE-A3-induced cytotoxic T lymphocytes using rAAV-6 capsid mutant vector.
abstract: MAGE-A3 is highly expressed in epithelial ovarian cancer (EOC), making it a promising candidate for immunotherapy. We investigated whether dendritic cells (DCs) transduced with a rAAV-6 capsid mutant vector Y445F could elicit effective MAGE-A3-specific anti-tumor cytotoxic T lymphocyte (CTL) responses in vitro. MAGE-A3 was cloned and rAAV-6-MAGE-A3 purified, followed by proviral genome detection using real-time PCR. Immunofluorescence detection of rAAV-6-Y445F-MAGE-A3-transduced DCs demonstrated 60% transduction efficiency. Fluorescent in situ hybridization analysis confirmed chromosomal integration of rAAV vectors. Flow cytometric analysis of transduced DCs showed unaltered expression of critical monocyte-derived surface molecules with retention of allo-stimulatory activity. Co-culture of autologous T lymphocytes with MAGE-A3-expressing DCs produced CTLs that secreted IFN-γ, and efficiently killed MAGE-A3+ EOC cells. This form of rAAV-based DC immunotherapy, either alone or more likely in combination with other immune-enhancing protocols, may prove useful in the clinical setting for management of EOC.
authors.affiliation: Laboratory of Surgical Oncology & Developmental Therapeutics, Department of Surgery, Wayne State University, Detroit, MI, USA; John D. Dingell VA Medical Center, Detroit, MI, USA; Virocan Therapeutics Pvt. Ltd., Guntur, Hydrabad, India. Electronic address: rbatchu@med.wayne.edu.
authors.forename: Ramesh B
authors.initials: RB
authors.lastname: Batchu
authors.affiliation: Laboratory of Surgical Oncology & Developmental Therapeutics, Department of Surgery, Wayne State University, Detroit, MI, USA; John D. Dingell VA Medical Center, Detroit, MI, USA.
authors.forename: Oksana V
authors.initials: OV
authors.lastname: Gruzdyn
authors.affiliation: Arkansas CLIA Lab, 401 South Cedar Street, Little Rock, AR, USA.
authors.forename: Amberly M
authors.initials: AM
authors.lastname: Moreno-Bost
authors.affiliation: Winthrop P. Rockefeller Cancer Institute, UAMS, Little Rock, AR, USA.
authors.forename: Susann
authors.initials: S
authors.lastname: Szmania
authors.affiliation: Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, FL, USA.
authors.forename: Giridhararao
authors.initials: G
authors.lastname: Jayandharan
authors.affiliation: Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, FL, USA.
authors.forename: Arun
authors.initials: A
authors.lastname: Srivastava
authors.affiliation: Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA.
authors.forename: Bala K
authors.initials: BK
authors.lastname: Kolli
authors.affiliation: Laboratory of Surgical Oncology & Developmental Therapeutics, Department of Surgery, Wayne State University, Detroit, MI, USA; Virocan Therapeutics Pvt. Ltd., Guntur, Hydrabad, India.
authors.forename: Donald W
authors.initials: DW
authors.lastname: Weaver
authors.affiliation: Winthrop P. Rockefeller Cancer Institute, UAMS, Little Rock, AR, USA.
authors.forename: Frits
authors.initials: F
authors.lastname: van Rhee
authors.affiliation: Laboratory of Surgical Oncology & Developmental Therapeutics, Department of Surgery, Wayne State University, Detroit, MI, USA; John D. Dingell VA Medical Center, Detroit, MI, USA.
authors.forename: Scott A
authors.initials: SA
authors.lastname: Gruber
mesh_terms: D000951:Antigens, Neoplasm / Q000276:immunology*; D002213:Capsid; D000077216:Carcinoma, Ovarian Epithelial; D045744:Cell Line, Tumor; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology; D000229:Dependovirus / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D007167:Immunotherapy; D007371:Interferon-gamma / Q000276:immunology; D007959:Lymphocyte Culture Test, Mixed; D009154:Mutation; D009363:Neoplasm Proteins / Q000276:immunology*; D009375:Neoplasms, Glandular and Epithelial / Q000473:pathology*; D010051:Ovarian Neoplasms / Q000473:pathology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014161:Transduction, Genetic
chemical_list: D000951:Antigens, Neoplasm; C072013:MAGEA3 protein, human; D009363:Neoplasm Proteins; D007371:Interferon-gamma
keywords: Adeno-associated Virus 6; Capsid tyrosine mutant; Cytotoxic T lymphocytes; Dendritic cells; Epithelial ovarian cancer; MAGE-A3