Immune Netw.
2006 Dec;6(4):192-198. 10.4110/in.2006.6.4.192.
Enhancement of Adenoviral Transduction and Immunogenecity of Transgenes by Soluble Coxsackie and Adenovirus Receptor-TAT Fusion Protein on Dendritic Cells
- Affiliations
-
- 1Department of Microbiology and Immunology, College of Medicine, The Catholic University of Korea. kimtg@catholic.ac.kr
- 2Catholic Hematopoietic Stem Cell Bank, The Catholic University of Korea.
- 3Department of Surgery, Kangnam St. Mary's Hospital, College of Medicine, The Catholic University of Korea.
- 4Boryung Biopharma Co. Ltd., Seoul, Korea.
- KMID: 1513151
- DOI: http://doi.org/10.4110/in.2006.6.4.192
Abstract
- BACKGROUND
Investigating strategy to enhance efficiency of gene transfer via adenovirus is critical to sustain gene expression in targeted cells or tissues to regulate immune responses. However, the use of adenovirus as a gene delivery method has been limited by the native tropism of the virus. In this study, the critical parameter is to improve the efficient binding of viral particles to the plasma membrane prior to cellular uptake.
METHODS
Human immunodeficiency virus (HIV-1) trans-acting activator of transcription (TAT), a protein transduction domain, was fused to the ectodomain of the coxsackie-adenovirus receptor (CAR). The CAR-TAT protein was produced from a Drosophila Schneider 2 cells (S2) transfected with CAR-TAT genes. The function of CAR- TAT was analyzed the efficiency of adenoviral gene transfer by flow cytometry, and then immunizing AdVGFP with CAR-TAT was transduced on dendritic cells (DCs).
RESULTS
S2 transfectants secreting CAR-TAT fusion protein has been stable over a period of 6 months and its expression was verified by western blot. Addition of CAR-TAT induced higher transduction efficiency for AdVGFP at every MOI tested. When mice were vaccinated with DC of which adenoviral transduction was mediated by CAR-TAT, the number of IFN-gamma secreting T-cells was increased as compared with those DCs transduced without CAR-TAT.
CONCLUSION
Our data provide evidence that CAR-TAT fusion protein enhances adenoviral transduction and immunogenecity of transgenes on DCs and may influence on the development of adenoviral- mediated anti-tumor immunotherapy.
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