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Short‐term culture of myeloid leukemic cells allows efficient transduction by adenoviral vectors
Author(s) -
Vereecque Rodolphe,
Saudemont Aurore,
Quesnel Bruno
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.568
Subject(s) - transduction (biophysics) , multiplicity of infection , cell culture , biology , viral vector , ex vivo , microbiology and biotechnology , genetic enhancement , transgene , myeloid , cancer research , in vivo , gene , recombinant dna , genetics , biochemistry
Background Ex vivo gene therapy of acute myeloid leukemia (AML) requires efficient transduction of leukemic cells. Recombinant adenovirus has been reported to be a poorly efficient vector in leukemic cells. We investigated leukemic cell culture as a possible method of improving the efficacy of this vector. Methods Leukemic cell lines and primary cultured AML cells were incubated with adenoviral vectors carrying GFP, LacZ, or IL‐12 cDNA. Transduction efficiency was evaluated by measuring adenoviral genome copy number and transgene expression in leukemic cells. The expression of the coxsackie/adenovirus receptor (CAR), CD29, CD49e, and CD51/61 was measured, as was the effect of blocking integrin on adenoviral transduction. Results Increasing the multiplicity of infection (MOI) to 300 plaque‐forming units per cell enhanced transduction of leukemic cell lines and to a lesser degree of AML cells. Analysis of adenoviral genome copy per cell showed only a partial correlation between gene transfer efficiency and transgene expression. Culture of AML cells for 3 days prior to adenoviral transduction increased both adenoviral copy number per cell and the percentage of transgene‐expressing cells. CD29, CD49e, and CD51/61 but not CAR expression increased in cultured AML cells between days 0 and 3 and integrin‐blocking experiments showed inhibition of transduction in two of four AML samples tested. Conclusions Efficient ex vivo gene transfer in primary cultured AML cells can be achieved by short‐term culture of leukemic cells prior to gene transfer with adenoviral vectors at a high MOI. This effect appears to be at least partially mediated by enhanced integrin expression. Copyright © 2004 John Wiley & Sons, Ltd.