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

S. Barrette, Janet Douglas, N. E. Seidel, D. M. Bodine

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

The low levels of transduction of human hematopoietic stem cells (HSCs) with Moloney murine leukemia virus (MLV) vectors have been an obstacle to gene therapy for hematopoietic diseases. It has been demonstrated that lentivirus vectors are more efficient than MLV vectors at transducing nondividing cell lines as well as human CD34+ cells and severe combined immunodeficiency disease re-populating cells. We compared transduction of cell lines and Lin- bone marrow cells, using a vesicular stomatitis virus G (VSV-G)-pseudotyped lentivirus or MLV vectors carrying a green fluorescent protein marker gene. As predicted, the lentivirus vector was more efficient at transducing mouse and human growth-inhibited cell lines. The transduction of mouse HSC by lentivirus vectors was compared directly to MLV vectors in a co-transduction assay. In this assay, transduction by ecotropic MLV is a positive internal control for downstream steps in retrovirus transduction, including cell division. Both the VSV-G lentivirus and MLV vectors transduced mouse HSCs maintained in cytokine-free medium at very low frequency, as did the ecotropic control. The lentivirus vector and the MLV vector were equally efficient at transducing bone marrow HSCs cultured in interleukin 3 (IL-3), IL-6, and stem cell factor for 96 hours. In conclusion, although lentivirus vectors are able to transduce growth-inhibited cell lines, the cell cycle status 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 The American Society of Hematology.

Original languageEnglish (US)
Pages (from-to)3385-3391
Number of pages7
JournalBlood
Volume96
Issue number10
StatePublished - Nov 15 2000
Externally publishedYes

Fingerprint

Moloney murine leukemia virus
Lentivirus
Hematopoietic Stem Cells
Stem cells
Viruses
Murine Leukemia Viruses
Cells
Cell Line
Vesicular Stomatitis
Cell growth
Assays
Bone
Severe Combined Immunodeficiency
Stem Cell Factor
Interleukin-3
Retroviridae
Gene therapy
Growth
Green Fluorescent Proteins
Bone Marrow Cells

ASJC Scopus subject areas

  • Hematology

Cite this

Lentivirus-based vectors transduce mouse hematopoietic stem cells with similar efficiency to Moloney murine leukemia virus-based vectors. / Barrette, S.; Douglas, Janet; Seidel, N. E.; Bodine, D. M.

In: Blood, Vol. 96, No. 10, 15.11.2000, p. 3385-3391.

Research output: Contribution to journalArticle

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