Lentivirus-based vectors transduce mouse hematopoietic stem cells with similar efficiency to Moloney murine leukemia virus-based vectors

Citation
S. Barrette et al., Lentivirus-based vectors transduce mouse hematopoietic stem cells with similar efficiency to Moloney murine leukemia virus-based vectors, BLOOD, 96(10), 2000, pp. 3385-3391
Citations number
63
Categorie Soggetti
Hematology,"Cardiovascular & Hematology Research
Journal title
BLOOD
ISSN journal
00064971 → ACNP
Volume
96
Issue
10
Year of publication
2000
Pages
3385 - 3391
Database
ISI
SICI code
0006-4971(20001115)96:10<3385:LVTMHS>2.0.ZU;2-2
Abstract
The low levels of transduction of human hematopoietic stem cells (HSCs) wit h Moloney murine leukemia virus (MLV) vectors have been an obstacle to gene therapy for hematopoietic diseases. It has been demonstrated that lentivir us vectors are more efficient than MLV vectors at transducing nondividing c ell lines as well as human CD34(+) cells and severe combined immunodeficien cy disease repopulating cells. We compared transduction of cell lines and L in(-) bone marrow cells, using a vesicular stomatitis virus G (VSV-G)-pseud otyped lentivirus or MLV vectors carrying a green fluorescent protein marke r gene. As predicted, the lentivirus vector was more efficient at transduci ng mouse and human growth-inhibited cell lines. The transduction of mouse H SC by lentivirus vectors was compared directly to MLV vectors in a co-trans duction assay. In this assay, transduction by ecotropic MLV is a positive i nternal control for downstream steps in retrovirus transduction, including cell division. Both the VSV-G lentivirus and MLV vectors transduced mouse H SCs maintained in cytokine-free medium at very low frequency, as did the ec otropic control. The lentivirus vector and the MLV vector were equally effi cient at transducing bone marrow HSCs cultured in interleukin 3 (IL-3), IL- 6, and stem cell factor for 96 hours. In conclusion, although lentivirus ve ctors are able to transduce growth-inhibited cell lines, the cell cycle sta tus of HSCs render them resistant to lentivirus-mediated transduction, and it is hypothesized that entry into cycle, not necessarily division, may be a requirement for efficient lentivirus-mediated transduction. (C) 2000 by T he American Society of Hematology.