627. Calcium Phopspate Precipitation of Adenovirus Enhances Gene Delivery into Human Dendritic Cells
Year: 2006
Type: article
Abstract: Top of pageAbstract
Mature dendritic cells (DC) are potent antigen presenting cells (APC) that have been used in vaccine studies as well as adoptive immunotherapy protocols. Adenovirus (Ad) vectors are frequently used to genetically modify DC; however gene transfer with recombinant adenovirus (serotype 5) into human DC is inefficient since DC do not express the cognate Ad5 receptor. Calcium phosphate (CaPi) precipitation of Ad has been shown to enhance gene transfer into murine DC and the aim of this study was to evaluate if CaPi precipitation enhances the gene delivery into human DC without interfering with their ability to activate antigen-specific T cells. Immature, monocyte derived DC were transduced with Ad5 or Ad5:CaPi complexes and gene expression was determined using GFP as a marker gene. CaPi precipitation of Ad5 increased DC gene transfer on average 35-fold when compared to unmodified Ad5. Ad5:CaPi complexes mediated gene delivery was similar in direct comparison to the optimal Ad for DC gene transfer (Ad5f35). Moreover, CaPi also increased the transduction efficiency of Ad5f35 at least 2-fold. CaPi transduction of immature DC did not reduce cell viability or their ability to be matured as judged by expression of CD83, and DR. To determine if Ad:CaPi treated DC can reactivate antigen-specific T cells, DC were incubated with an Ad5f35 vector encoding the subdominant Epstein Barr virus (EBV) antigens, latent membrane proteins (LMP) 2, which is expressed in EBV-positive malignancies. CaPi precipitation increased the expression of LMP2 in human DC 5|[ndash]|10 fold compared to virus alone, resulting in enhanced activation of LMP2-specific T cells in vitro. In conclusion, CaPi complexes increased gene transfer into human DC with both Ad5 and Ad5f35 vectors without changing their phenotype or their ability to activate antigen-specific T cells. Ad5:CaPi mediated transduction resulted in similar transduction as Ad5f35 vectors, which allows the use of existing Ad5 vector technology to genetically modify human DC.
Source: Molecular Therapy
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $4,500
OpenAlex ID: W2069533949