Efficient lysis of epithelial ovarian cancer cells by MAGE-A3-induced cytotoxic T lymphocytes using rAAV-6 capsid mutant vector

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• 作者：Ramesh B. Batchu ; Oksana V. Gruzdyn ; Amberly M. Moreno-Bost ; Susann Szmania ; Giridhararao Jay ; haran ; Arun Srivastava ; Bala K. Kolli ; Donald W. Weaver ; Frits van Rhee ; Scott A. Gruber
• 关键词：Epithelial ovarian cancer ; MAGE-A3 ; Adeno-associated Virus 6 ; Capsid tyrosine mutant ; Dendritic cells ; Cytotoxic T lymphocytes
• 刊名：Vaccine
• 出版年：12 February, 2014
• 年：2014
• 卷：32
• 期：8
• 页码：938-943
• 全文大小：1263 K

文摘

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-纬, 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.