Development of an adenoviral vector system with adenovirus serotype 35 tropism; efficient transient gene transfer into primary malignant hematopoietic cells

✍ Scribed by Marcus Nilsson; Johan Ljungberg; Johan Richter; Thomas Kiefer; Mattias Magnusson; André Lieber; Bengt Widegren; Stefan Karlsson; Xiaolong Fan

• Publisher: John Wiley and Sons
• Year: 2004
• Tongue: English
• Weight: 245 KB
• Volume: 6
• Category: Article
• ISSN: 1099-498X
• DOI: 10.1002/jgm.543

✦ Synopsis

Abstract

Background

A paucity of coxsackie adenovirus receptor (CAR) hampers the adenovirus serotype 5 (Ad5)‐based vector‐mediated gene transfer into malignant hematopoietic cells. Fiber‐retargeted adenoviral vectors with species B tropism can potentially bypass the CAR requirement and facilitate efficient gene transfer into malignant hematopoietic cells.

Methods

For feasible generation of fiber‐retargeted adenoviral vectors, we have modified the versatile AdEasy system with a chimeric fiber gene encoding the Ad5 fiber tail domain and Ad35 fiber shaft and knob domains. An Ad5‐based vector encoding the green fluorescent protein (GFP) gene under the control of the PGK promoter with Ad35 fiber receptor specificity was generated (Ad5F35‐GFP). The Ad5F35‐GFP vector‐mediated gene transfer efficiency was compared with a fiber non‐modified Ad5‐GFP vector, which also encodes the GFP gene under the control of the PGK promoter.

Results

We demonstrated that a variety of Ad5‐refractory malignant myeloid and B lymphoid cell lines were highly permissive to the Ad5F35‐GFP vector infection. Importantly, primary chronic myeloid leukemic (CML) cells and chronic lymphocytic leukemia (CLL) B cells were superiorly transduced by the Ad5F35‐GFP vector at a multiplicity of infection (MOI) of 100 compared with the Ad5‐GFP vector.

Conclusions

Our study will facilitate the generation of fiber‐retargeted adenoviral vectors and enable transient genetic manipulation of primary malignant hematopoietic cells. Copyright © 2004 John Wiley & Sons, Ltd.