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Enhancer‐deleted retroviral vectors restore high levels of superoxide generation in a mouse model of CGD
Author(s) -
Schwickerath Oliver,
Brouns Gaby,
Thrasher Adrian,
Kin Christine,
Roes Jürgen,
Casimir Colin
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.557
Subject(s) - enhancer , transduction (biophysics) , viral vector , genetic enhancement , haematopoiesis , biology , long terminal repeat , bone marrow , vector (molecular biology) , microbiology and biotechnology , gene , myeloid , transfection , virology , stem cell , immunology , genetics , gene expression , recombinant dna , biochemistry
Background Retroviral vectors possess many advantages for use in gene therapy protocols, especially within the haematopoietic system. A number of attendant problems, however, still limit their safety in clinical application. The effects of the enhancer present in the retroviral long terminal repeat (LTR) are a major concern for the clinical usage of such vectors, as they can exert a powerful regulatory influence on the genes that surround them. Methods To improve the safety and widen the applicability of retroviral vectors for use in gene therapy we have developed an enhancer‐deleted (Δ‐LTR) retroviral vector that retained high titre and demonstrated transcriptional activity in myeloid cells. Results When used to correct a mouse model of autosomal recessive chronic granulomatous disease, the Δ‐LTR vectors gave acceptable levels of gene transfer to mouse bone marrow cells. Evidence for a slight preferential expression in myeloid cells was obtained with all the vectors studied. Nitroblue tetrazolium assay of superoxide generation in mouse bone marrow derived haematopoietic colonies revealed that transduction with Δ‐LTR vectors could restore functional NADPH oxidase to cells from these animals. Superoxide assay of peripheral blood confirmed that, although relatively low numbers of cells were transduced, the Δ‐LTR vector was capable of reconstituting very high levels of oxidase activity, comparable to that obtained from normal cells. Conclusions The Δ‐LTR vector described here could provide the basis for a new generation of retroviral vectors with improved safety. Copyright © 2004 John Wiley & Sons, Ltd.