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

George V. Aslanidi, Angela E. Rivers, Luis Ortiz, Lakshmanan Govindasamy, Chen Ling, Giridhara R. Jayandharan, Sergei Zolotukhin, Mavis Agbandje-McKenna, Arun Srivastava

Research output: Contribution to journalArticle

32 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)3908-3917
Number of pages10
JournalVaccine
Volume30
Issue number26
DOIs
StatePublished - Jun 6 2012
Externally publishedYes

Keywords

  • Adeno-associated virus vectors
  • Capsid proteins
  • Dendritic cells
  • Gene expression
  • Serine-phosphorylation
  • Serine/threonine kinase

Fingerprint Dive into the research topics of 'High-efficiency transduction of human monocyte-derived dendritic cells by capsid-modified recombinant AAV2 vectors'. Together they form a unique fingerprint.

Cite this