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Efficient transduction of pigtailed macaque hematopoietic repopulating cells with HIV-based lentiviral vectors.

Abstract：

:Lentiviral vectors are attractive for hematopoietic stem cell (HSC) gene therapy because they do not require mitosis for nuclear entry, they efficiently transduce hematopoietic repopulating cells, and self-inactivating (SIN) designs can be produced at high titer. Experiments to evaluate HIV-derived lentiviral vectors in nonhuman primates prior to clinical trials have been hampered by low transduction frequencies due in part to host restriction by TRIM5alpha. We have established conditions for efficient transduction of pigtailed macaque (Macaca nemestrina) long-term repopulating cells using VSV-G-pseudotyped HIV-based lentiviral vectors. Stable, long-term, high-level gene marking was observed in 3 macaques using relatively low MOIs (5-10) in a 48-hour ex vivo transduction protocol. All animals studied had rapid neutrophil engraftment with a median of 10.3 days to a count greater than 0.5 x 10(9)/L (500/microL). Expression was detected in all lineages, with long-term marking levels in granulocytes at approximately 20% to 30%, and in lymphocytes at approximately 12% to 23%. All animals had polyclonal engraftment as determined by analysis of vector integration sites. These data suggest that lentiviral vectors should be highly effective for HSC gene therapy, particularly for diseases in which maintaining the engraftment potential of stem cells using short-term ex vivo transduction protocols is critical.

journal_name

Blood

journal_title

Blood

authors

Trobridge GD,Beard BC,Gooch C,Wohlfahrt M,Olsen P,Fletcher J,Malik P,Kiem HP

doi

10.1182/blood-2007-09-115022

subject

Has Abstract

pub_date

2008-06-15 00:00:00

pages

5537-43

issue

12

eissn

0006-4971

issn

1528-0020

pii

blood-2007-09-115022

journal_volume

111

pub_type

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