High-efficiency transduction of human monocyte-derived dendritic cells by capsid-modified recombinant AAV2 vectors
| Autor: | Arun Srivastava, Sergei Zolotukhin, Giridhara R. Jayandharan, Mavis Agbandje-McKenna, Luis Ortiz, Lakshmanan Govindasamy, Chen Ling, Angela Rivers, George Aslanidi |
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| Rok vydání: | 2012 |
| Předmět: |
Genetic Vectors
Biology Article law.invention Serine Transduction (genetics) Transduction Genetic law Humans Cytotoxic T cell Telomerase reverse transcriptase Cells Cultured Serine/threonine-specific protein kinase General Veterinary General Immunology and Microbiology Adenoviruses Human Public Health Environmental and Occupational Health Dendritic Cells Molecular biology Infectious Diseases Amino Acid Substitution Mutagenesis Site-Directed Recombinant DNA Molecular Medicine Phosphorylation Capsid Proteins Mutant Proteins K562 cells |
| Zdroj: | Vaccine. 30:3908-3917 |
| ISSN: | 0264-410X |
| DOI: | 10.1016/j.vaccine.2012.03.079 |
| Popis: | Phosphorylation of surface-exposed tyrosine residues negatively impacts the transduction efficiency of recombinant AAV2 vectors. Pre-treatment of cells with specific cellular serine/threonine kinase inhibitors also significantly increased the transduction efficiency of AAV2 vectors. We reasoned that site-directed mutagenesis of surface-exposed serine residues might allow the vectors to evade phosphorylation and thus lead to higher transduction efficiency. Each of the 15 surface-exposed serine (S) residues was substituted with valine (V) residues, and the transduction efficiency of three of these mutants, S458V, S492V and S662V, was increased by up to ≈ 20-fold in different cell types. The S662V mutant was efficient in transducing human monocyte-derived dendritic cells (moDCs), a cell type not readily amenable to transduction by the conventional AAV vectors, and did not induce any phenotypic changes in these cells. Recombinant S662V-AAV2 vectors encoding a truncated human telomerase (hTERT) gene were generated and used to stimulate cytotoxic T cells (CTLs) against target cells. S662V-AAV2-hTERT vector-transduced DCs resulted in rapid, specific T-cell clone proliferation and generation of robust CTLs, which led to specific cell lysis of K562 cells. These studies suggest that high-efficiency transduction of moDCs by serine-modified AAV2 vectors is feasible, which supports the potential utility of these vectors for future human DCs vaccine studies. |
| Databáze: | OpenAIRE |
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