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High levels of transgene expression following transduction of long-term NOD/SCID-repopulating human cells with a modified lentiviral vector.
Stem Cells. 2001; 19(3):247-59.SC

Abstract

Both oncoretroviral and lentiviral vectors have been shown to transduce CD34(+) human hematopoietic stem cells (HSC) capable of establishing human hematopoiesis in nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice that support partially human hematopoiesis. We and others have reported that murine stem cell virus (MSCV)-based oncoretroviral vectors efficiently transduced HSC that had been cultured ex vivo for 4-7 days with cytokines, resulting in transgene expression in lymphoid and myeloid progenies of SCID-engrafting cells 4-8 weeks post-transplantation. Although lentiviral vectors have been demonstrated to transduce HSC under minimal ex vivo culture conditions, concerns exist regarding the level of transgene expression mediated by these vectors. We therefore evaluated a novel hybrid lentiviral vector (GIN-MU3), in which the U3 region of the HIV-1 long terminal repeat was replaced by the MSCV U3 region (MU3). Human cord blood CD34(+) cells were transduced with vesicular stomatitis virus G envelope protein-pseudotyped lentiviruses during a 48-hour culture period. After a total of 4 days in culture, transduced cells were transplanted into NOD/SCID mice to examine gene transfer and expression in engrafting human cells. Fifteen weeks post-transplantation, 37% +/- 12% of engrafted human cells expressed the green fluorescence protein (GFP) gene introduced by the lentiviral vector. High levels of GFP expression were observed in lymphoid, myeloid and erythroid progenies, and in engrafted human cells that retained the CD34(+) phenotype 15 weeks post-transplantation. This study provides evidence that lentiviral vectors transduced both short-term and long-term engrafting human cells, and mediated persistent transgene expression at high levels in multiple lineages of hematopoietic cells.

Authors+Show Affiliations

Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21231, USANo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

11359950

Citation

Gao, Z, et al. "High Levels of Transgene Expression Following Transduction of Long-term NOD/SCID-repopulating Human Cells With a Modified Lentiviral Vector." Stem Cells (Dayton, Ohio), vol. 19, no. 3, 2001, pp. 247-59.
Gao Z, Golob J, Tanavde VM, et al. High levels of transgene expression following transduction of long-term NOD/SCID-repopulating human cells with a modified lentiviral vector. Stem Cells. 2001;19(3):247-59.
Gao, Z., Golob, J., Tanavde, V. M., Civin, C. I., Hawley, R. G., & Cheng, L. (2001). High levels of transgene expression following transduction of long-term NOD/SCID-repopulating human cells with a modified lentiviral vector. Stem Cells (Dayton, Ohio), 19(3), 247-59.
Gao Z, et al. High Levels of Transgene Expression Following Transduction of Long-term NOD/SCID-repopulating Human Cells With a Modified Lentiviral Vector. Stem Cells. 2001;19(3):247-59. PubMed PMID: 11359950.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - High levels of transgene expression following transduction of long-term NOD/SCID-repopulating human cells with a modified lentiviral vector. AU - Gao,Z, AU - Golob,J, AU - Tanavde,V M, AU - Civin,C I, AU - Hawley,R G, AU - Cheng,L, PY - 2001/5/22/pubmed PY - 2001/7/28/medline PY - 2001/5/22/entrez SP - 247 EP - 59 JF - Stem cells (Dayton, Ohio) JO - Stem Cells VL - 19 IS - 3 N2 - Both oncoretroviral and lentiviral vectors have been shown to transduce CD34(+) human hematopoietic stem cells (HSC) capable of establishing human hematopoiesis in nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice that support partially human hematopoiesis. We and others have reported that murine stem cell virus (MSCV)-based oncoretroviral vectors efficiently transduced HSC that had been cultured ex vivo for 4-7 days with cytokines, resulting in transgene expression in lymphoid and myeloid progenies of SCID-engrafting cells 4-8 weeks post-transplantation. Although lentiviral vectors have been demonstrated to transduce HSC under minimal ex vivo culture conditions, concerns exist regarding the level of transgene expression mediated by these vectors. We therefore evaluated a novel hybrid lentiviral vector (GIN-MU3), in which the U3 region of the HIV-1 long terminal repeat was replaced by the MSCV U3 region (MU3). Human cord blood CD34(+) cells were transduced with vesicular stomatitis virus G envelope protein-pseudotyped lentiviruses during a 48-hour culture period. After a total of 4 days in culture, transduced cells were transplanted into NOD/SCID mice to examine gene transfer and expression in engrafting human cells. Fifteen weeks post-transplantation, 37% +/- 12% of engrafted human cells expressed the green fluorescence protein (GFP) gene introduced by the lentiviral vector. High levels of GFP expression were observed in lymphoid, myeloid and erythroid progenies, and in engrafted human cells that retained the CD34(+) phenotype 15 weeks post-transplantation. This study provides evidence that lentiviral vectors transduced both short-term and long-term engrafting human cells, and mediated persistent transgene expression at high levels in multiple lineages of hematopoietic cells. SN - 1066-5099 UR - https://www.unboundmedicine.com/medline/citation/11359950/High_levels_of_transgene_expression_following_transduction_of_long_term_NOD/SCID_repopulating_human_cells_with_a_modified_lentiviral_vector_ L2 - https://doi.org/10.1634/stemcells.19-3-247 DB - PRIME DP - Unbound Medicine ER -