Premium
Development of an adenoviral vector system with adenovirus serotype 35 tropism; efficient transient gene transfer into primary malignant hematopoietic cells
Author(s) -
Nilsson Marcus,
Ljungberg Johan,
Richter Johan,
Kiefer Thomas,
Magnusson Mattias,
Lieber André,
Widegren Bengt,
Karlsson Stefan,
Fan Xiaolong
Publication year - 2004
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.543
Subject(s) - biology , tropism , viral vector , green fluorescent protein , virology , vector (molecular biology) , multiplicity of infection , genetic enhancement , haematopoiesis , gene , microbiology and biotechnology , stem cell , virus , genetics , recombinant dna
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.