Rationally designed capsid and transgene cassette of AAV6 vectors for dendritic cell-based cancer immunotherapy

被引:41
作者
Pandya, Jheel [1 ]
Ortiz, Luis [2 ]
Ling, Chen [2 ]
Rivers, Angela E. [3 ]
Aslanidi, George [2 ]
机构
[1] Univ Florida, Dept Microbiol & Cell Sci, Coll Med, Gainesville, FL 32610 USA
[2] Univ Florida, Dept Pediat, Coll Med, Div Cellular & Mol Therapy, Gainesville, FL 32610 USA
[3] Univ Illinois, Dept Pediat, Div Hematol Oncol, Chicago, IL USA
关键词
adeno-associated virus vectors; CD11c promoter; dendritic cells; gene expression serine/threonine phosphorylation; HIGH-EFFICIENCY TRANSDUCTION; ADENOASSOCIATED VIRUS TYPE-2; SEROTYPE VECTORS; NEXT-GENERATION; IMMUNE-RESPONSE; IN-VITRO; GENE; BINDING; CONSTRUCTION; SUBSETS;
D O I
10.1038/icb.2013.74
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Dendritic cell (DC)-based immunotherapy has recently demonstrated a great potential for clinical applications; however, additional progress in the methods of tumor-specific antigen delivery to DCs is necessary for the further development of antitumor vaccines. To this end, a capsid-optimized adeno-associated virus serotype 6 (AAV6-T492V+S663V) vector was developed by site-directed mutagenesis of surface-exposed serine (S) and threonine (T) residues, which have a critical role in intracellular trafficking of AAV vectors. This double-mutant AAV6 vector had similar to 5-fold greater transduction efficiency in monocyte-derived DCs (moDCs) compared with wild-type (WT)-AAV6 vectors. The increase in the transduction efficiency correlated with the improved nuclear translocation of AAV6-T492V+S663V over that of the WT-AAV6 vector. Additional studies of the CD11c promoter identified critical regulatory elements that fit into the AAV expression cassette and drive EGFP expression in moDCs. Development of a chimeric promoter (chmCD11c) that contains functional modules of CD11c and a Simian virus (SV40) enhancer element dramatically increased the EGFP expression in moDCs. MoDCs transduced by the capsid-optimized AAV6 vector carrying human prostate-specific antigen (hPSA) driven by CBA (AAV6-T492V+S663V-CBA-hPSA) or chmCd11c (AAV6T492V+S663V-chmCD11c-hPSA) generated specific T-cell clone proliferation and superior cytotoxic T lymphocytes (CTLs) with higher killing capability against human prostate adenocarcinoma cells, LNCaP, compared with WT-AAV6 induced CTLs. Taken together, these studies suggest that optimization of capsid and promoter components of AAV vectors can be a useful approach for efficient targeting of moDCs and may prove to be a promising tool for cancer immunotherapy.
引用
收藏
页码:116 / 123
页数:8
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