Stable transduction of quiescent CD34+CD38- human hematopoietic cells by HIV-1-based lentiviral vectors

Scott S. Case, Mary A. Price, Craig T. Jordan, Xiao Jin Yu, Lijun Wang, Gerhard Bauer, Dennis L. Haas, Dakun Xu, Renata Stripecke, Luigi Naldini, Donald B. Kohn, Gay M. Crooks

Research output: Contribution to journalArticle

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Abstract

We compared the efficiency of transduction by an HIV-1-based lentiviral vector to that by a Moloney murine leukemia virus (MLV) retroviral vector, using stringent in vitro assays of primitive, quiescent human hematopoietic progenitor cells. Each construct contained the enhanced green fluorescent protein (GFP) as a reporter gene. The lentiviral vector, but not the MLV vector, expressed GFP in nondivided CD34+ cells (45.5% GFP+) and in CD34+CD38- cells in G0 (12.4% GFP+), 48 hr after transduction. However, GFP could also be detected short-term in CD34+ cells transduced with a lentiviral vector that contained a mutated integrase gene. The level of stable transduction from integrated vector was determined after extended long-term bone marrow culture. Both MLV vectors and lentiviral vectors efficiently transduced cytokine-stimulated CD34+ cells. The MLV vector did not transduce more primitive, quiescent CD34+CD38- cells (n = 8). In contrast, stable transduction of CD34+CD38- cells by the lentiviral vector was seen for over 15 weeks of extended long-term culture (9.2 ± 5.2%, n = 7). GFP expression in clones from single CD34+CD38- cells confirmed efficient, stable lentiviral transduction in 29% of early and late- proliferating cells. In the absence of growth factors during transduction, only the lentiviral vector was able to transduce CD34+ and CD34+CD38- cells (13.5 ± 2.5%, n = 11 and 12.2 ± 9.7%, n = 4, respectively). The lentiviral vector is clearly superior to the MLV vector for transduction of quiescent, primitive human hematopoietic progenitor cells and may provide therapeutically useful levels of gene transfer into human hematopoietic stem cells.

Original languageEnglish (US)
Pages (from-to)2988-2993
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number6
DOIs
StatePublished - Mar 16 1999
Externally publishedYes

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HIV-1
Green Fluorescent Proteins
Murine Leukemia Viruses
Hematopoietic Stem Cells
Moloney murine leukemia virus
Integrases
Reporter Genes
Genes
Intercellular Signaling Peptides and Proteins
Clone Cells
Bone Marrow
Cytokines

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Stable transduction of quiescent CD34+CD38- human hematopoietic cells by HIV-1-based lentiviral vectors. / Case, Scott S.; Price, Mary A.; Jordan, Craig T.; Yu, Xiao Jin; Wang, Lijun; Bauer, Gerhard; Haas, Dennis L.; Xu, Dakun; Stripecke, Renata; Naldini, Luigi; Kohn, Donald B.; Crooks, Gay M.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 96, No. 6, 16.03.1999, p. 2988-2993.

Research output: Contribution to journalArticle

Case, SS, Price, MA, Jordan, CT, Yu, XJ, Wang, L, Bauer, G, Haas, DL, Xu, D, Stripecke, R, Naldini, L, Kohn, DB & Crooks, GM 1999, 'Stable transduction of quiescent CD34+CD38- human hematopoietic cells by HIV-1-based lentiviral vectors', Proceedings of the National Academy of Sciences of the United States of America, vol. 96, no. 6, pp. 2988-2993. https://doi.org/10.1073/pnas.96.6.2988
Case, Scott S. ; Price, Mary A. ; Jordan, Craig T. ; Yu, Xiao Jin ; Wang, Lijun ; Bauer, Gerhard ; Haas, Dennis L. ; Xu, Dakun ; Stripecke, Renata ; Naldini, Luigi ; Kohn, Donald B. ; Crooks, Gay M. / Stable transduction of quiescent CD34+CD38- human hematopoietic cells by HIV-1-based lentiviral vectors. In: Proceedings of the National Academy of Sciences of the United States of America. 1999 ; Vol. 96, No. 6. pp. 2988-2993.
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abstract = "We compared the efficiency of transduction by an HIV-1-based lentiviral vector to that by a Moloney murine leukemia virus (MLV) retroviral vector, using stringent in vitro assays of primitive, quiescent human hematopoietic progenitor cells. Each construct contained the enhanced green fluorescent protein (GFP) as a reporter gene. The lentiviral vector, but not the MLV vector, expressed GFP in nondivided CD34+ cells (45.5{\%} GFP+) and in CD34+CD38- cells in G0 (12.4{\%} GFP+), 48 hr after transduction. However, GFP could also be detected short-term in CD34+ cells transduced with a lentiviral vector that contained a mutated integrase gene. The level of stable transduction from integrated vector was determined after extended long-term bone marrow culture. Both MLV vectors and lentiviral vectors efficiently transduced cytokine-stimulated CD34+ cells. The MLV vector did not transduce more primitive, quiescent CD34+CD38- cells (n = 8). In contrast, stable transduction of CD34+CD38- cells by the lentiviral vector was seen for over 15 weeks of extended long-term culture (9.2 ± 5.2{\%}, n = 7). GFP expression in clones from single CD34+CD38- cells confirmed efficient, stable lentiviral transduction in 29{\%} of early and late- proliferating cells. In the absence of growth factors during transduction, only the lentiviral vector was able to transduce CD34+ and CD34+CD38- cells (13.5 ± 2.5{\%}, n = 11 and 12.2 ± 9.7{\%}, n = 4, respectively). The lentiviral vector is clearly superior to the MLV vector for transduction of quiescent, primitive human hematopoietic progenitor cells and may provide therapeutically useful levels of gene transfer into human hematopoietic stem cells.",
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AU - Wang, Lijun

AU - Bauer, Gerhard

AU - Haas, Dennis L.

AU - Xu, Dakun

AU - Stripecke, Renata

AU - Naldini, Luigi

AU - Kohn, Donald B.

AU - Crooks, Gay M.

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