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 Blood, 1 July 2005, Vol. 106, No. 1, pp. 51-58.
Prepublished online as a Blood First Edition Paper on March 17, 2005; DOI 10.1182/blood-2004-11-4491.

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Submitted November 29, 2004
Accepted March 8, 2005

Improved transduction of human sheep repopulating cells by retrovirus vectors pseudotyped with feline leukemia virus type C or RD114 envelopes

M L Lucas, Nancy E Seidel, Christopher D Porada, John G Quigley, Stacie M Anderson, Harry L Malech, Janis L Abkowitz, Esmail D Zanjani, and David M Bodine*

GMBB, NHGRI/NIH, Bethesda, MD, USA
University of Nevada, Reno, NV, USA
University of Washington, Seattle, WA, USA
NIAID/NIH, Bethesda, MD, USA

* Corresponding author; email: tedyaz{at}mail.nih.gov.

Gene therapy for hematopoietic diseases has been hampered by the low frequency of transduction of human hematopoietic stem cells (HSC) with retroviral vectors pseudotyped with amphotropic envelopes. We hypothesized that transduction could be increased by the use of retroviral vectors pseudotyped with envelopes that recognize more abundant cellular receptors. The levels of mRNA encoding the receptors of the feline retroviruses, RD114 and Feline Leukemia Virus Type-C (FeLV-C), were significantly higher than the level of Gibbon Ape Leukemia Virus (GaLV) receptor mRNA in cells enriched for human HSC (Lin- CD34+ CD38-). We co-transduced human peripheral blood CD34+ cells with equivalent numbers of FeLV-C and GALV or RD114 and GaLV pseudotyped retroviruses for injection into fetal sheep. Analysis of DNA from peripheral blood and bone marrow from recipient sheep demonstrated that FeLV-C or RD114 pseudotyped vectors were present at significantly higher levels than GALV pseudotyped vectors. Analysis of individual myeloid colonies demonstrated that retrovirus vectors with FeLV-C and RD114 pseudotypes were present at 1.5-1.6 copies per cell and were preferentially integrated near known genes. We conclude that the more efficient transduction of human HSC with either FeLV-C or RD114 pseudotyped retroviral particles may improve gene transfer in human clinical trials.


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Improved human stem-cell transduction: the cat's meow
Brian P. Sorrentino
Blood 2005 106: 8. [Full Text] [PDF]





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