| Short-term culture of myeloid leukemic cells allows efficient transduction by adenoviral vectors. | |
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MedLine Citation:
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PMID: 15241782 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Rodolphe Vereecque; Aurore Saudemont; Bruno Quesnel |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: The journal of gene medicine Volume: 6 ISSN: 1099-498X ISO Abbreviation: J Gene Med Publication Date: 2004 Jul |
Date Detail:
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Created Date: 2004-07-08 Completed Date: 2005-02-23 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 9815764 Medline TA: J Gene Med Country: England |
Other Details:
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Languages: eng Pagination: 751-9 Citation Subset: IM |
Copyright Information:
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Copyright 2004 John Wiley & Sons, Ltd. |
Affiliation:
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Unité INSERM 524, Institut de Recherche sur le Cancer de Lille, IFR-114, France. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Acute Disease Adenoviridae / genetics* Cell Line, Tumor Flow Cytometry Genetic Vectors* Genome, Viral Humans Leukemia, Myeloid / genetics, pathology* Polymerase Chain Reaction Transduction, Genetic* |
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