High-efficiency transduction of human monocyte-derived dendritic cells by capsid-modified recombinant AAV2 vectors

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• 作者：George V. Aslanidi^a ; ^b ; ^{aslanidi@peds.ufl.edu} ; Angela E. Rivers^a ; Luis Ortiz^a ; Lakshmanan Govindasamy^c ; Chen Ling^a ; ^b ; ^d ; Giridhara R. Jayandharan^a ; ^b ; ^g ; Sergei Zolotukhin^a ; ^b ; ^d ; ^e ; Mavis Agbandje-McKenna^b ; ^c ; ^e ; Arun Srivastava^a ; ^b ; ^d ; ^e ; ^f ; ^{aruns@peds.ufl.edu}
• 关键词：Adeno-associated virus vectors ; Capsid proteins ; Serine-phosphorylation ; Serine/threonine kinase ; Dendritic cells ; Gene expression
• 出版年：2012
• 期刊代码：89_0264410x
• 类别：imm
• 出版时间：6 June, 2012
• 卷：30
• 期：26
• 页码：3908-3917
• 文件大小：1605 K

摘要

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 鈭?0-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.