Lentiviral gene transfer into primary and secondary NOD/SCID repopulating cells
Woods, N B LU ; Fahlman, C ; Mikkola, H ; Hamaguchi, I LU ; Olsson, K LU ; Zufferey, R ; Jacobsen, S E LU ; Trono, D and Karlsson, S LU
(2000)
In Blood
96(12).
p.33-3725
- Abstract
The ability of lentiviral vectors to transfer genes into human hematopoietic stem cells was studied, using a human immunodeficiency virus 1 (HIV-1)-derived vector expressing the green fluorescence protein (GFP) downstream of the phosphoglycerate kinase (PGK) promoter and pseudotyped with the G protein of vesicular stomatitis virus (VSV). High-efficiency transduction of human cord blood CD34(+) cells was achieved after overnight incubation with vector particles. Sixteen to 28 percent of individual colony-forming units granulocyte-macrophage (CFU-GM) colonies derived from cord blood CD34(+) cells were positive by polymerase chain reaction (PCR) for the GFP gene. The transduction efficiency of SCID-repopulating cells (SRC) within the cord... (More)
The ability of lentiviral vectors to transfer genes into human hematopoietic stem cells was studied, using a human immunodeficiency virus 1 (HIV-1)-derived vector expressing the green fluorescence protein (GFP) downstream of the phosphoglycerate kinase (PGK) promoter and pseudotyped with the G protein of vesicular stomatitis virus (VSV). High-efficiency transduction of human cord blood CD34(+) cells was achieved after overnight incubation with vector particles. Sixteen to 28 percent of individual colony-forming units granulocyte-macrophage (CFU-GM) colonies derived from cord blood CD34(+) cells were positive by polymerase chain reaction (PCR) for the GFP gene. The transduction efficiency of SCID-repopulating cells (SRC) within the cord blood CD34(+) population was assessed by serial transplantation into nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. When 400,000 cord blood CD34(+) cells were transplanted into primary recipients, all primary and secondary recipients contained and expressed the transgene. Over 50% of CFU-GM colonies derived from the bone marrow of these primary and secondary recipients contained the vector on average as determined by PCR. Transplantation of transduced cells in limiting dilution generated GFP(+) lymphoid and myeloid progeny cells that may have arisen from a single SRC. Inverse PCR analysis was used to amplify vector-chromosomal junctional fragments in colonies derived from SRC and confirmed that the vector was integrated. These results show that lentiviral vectors can efficiently transduce very primitive human hematopoietic progenitor and stem cells. (Blood. 2000;96:3725-3733)
(Less)
- author
- Woods, N B
LU
; Fahlman, C
; Mikkola, H
; Hamaguchi, I
LU
; Olsson, K
LU
; Zufferey, R
; Jacobsen, S E
LU
; Trono, D
and Karlsson, S
LU
- organization
- publishing date
- 2000-12-01
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Animals, Antigens, CD34/physiology, Cell Lineage, Fetal Blood/cytology, Gene Transfer Techniques, Genetic Vectors/blood, Graft Survival, HIV-1/genetics, Hematopoiesis, Hematopoietic Stem Cell Transplantation, Hematopoietic Stem Cells/metabolism, Humans, Immunophenotyping, Lentivirus/genetics, Mice, Mice, Inbred NOD/blood, Mice, SCID/blood, Polymerase Chain Reaction, Transduction, Genetic/standards
- in
- Blood
- volume
- 96
- issue
- 12
- pages
- 33 - 3725
- publisher
- American Society of Hematology
- external identifiers
-
- pmid:11090053
- scopus:0034548823
- ISSN
- 0006-4971
- DOI
- 10.1182/blood.V96.12.3725.h8003725_3725_3733
- language
- English
- LU publication?
- yes
- id
- 40f674e7-3fb2-4689-8187-81993d82c265
- date added to LUP
- 2019-11-25 13:45:03
- date last changed
- 2024-10-02 17:08:01
@article{40f674e7-3fb2-4689-8187-81993d82c265,
abstract = {{<p>The ability of lentiviral vectors to transfer genes into human hematopoietic stem cells was studied, using a human immunodeficiency virus 1 (HIV-1)-derived vector expressing the green fluorescence protein (GFP) downstream of the phosphoglycerate kinase (PGK) promoter and pseudotyped with the G protein of vesicular stomatitis virus (VSV). High-efficiency transduction of human cord blood CD34(+) cells was achieved after overnight incubation with vector particles. Sixteen to 28 percent of individual colony-forming units granulocyte-macrophage (CFU-GM) colonies derived from cord blood CD34(+) cells were positive by polymerase chain reaction (PCR) for the GFP gene. The transduction efficiency of SCID-repopulating cells (SRC) within the cord blood CD34(+) population was assessed by serial transplantation into nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. When 400,000 cord blood CD34(+) cells were transplanted into primary recipients, all primary and secondary recipients contained and expressed the transgene. Over 50% of CFU-GM colonies derived from the bone marrow of these primary and secondary recipients contained the vector on average as determined by PCR. Transplantation of transduced cells in limiting dilution generated GFP(+) lymphoid and myeloid progeny cells that may have arisen from a single SRC. Inverse PCR analysis was used to amplify vector-chromosomal junctional fragments in colonies derived from SRC and confirmed that the vector was integrated. These results show that lentiviral vectors can efficiently transduce very primitive human hematopoietic progenitor and stem cells. (Blood. 2000;96:3725-3733)</p>}},
author = {{Woods, N B and Fahlman, C and Mikkola, H and Hamaguchi, I and Olsson, K and Zufferey, R and Jacobsen, S E and Trono, D and Karlsson, S}},
issn = {{0006-4971}},
keywords = {{Animals; Antigens, CD34/physiology; Cell Lineage; Fetal Blood/cytology; Gene Transfer Techniques; Genetic Vectors/blood; Graft Survival; HIV-1/genetics; Hematopoiesis; Hematopoietic Stem Cell Transplantation; Hematopoietic Stem Cells/metabolism; Humans; Immunophenotyping; Lentivirus/genetics; Mice; Mice, Inbred NOD/blood; Mice, SCID/blood; Polymerase Chain Reaction; Transduction, Genetic/standards}},
language = {{eng}},
month = {{12}},
number = {{12}},
pages = {{33--3725}},
publisher = {{American Society of Hematology}},
series = {{Blood}},
title = {{Lentiviral gene transfer into primary and secondary NOD/SCID repopulating cells}},
url = {{http://dx.doi.org/10.1182/blood.V96.12.3725.h8003725_3725_3733}},
doi = {{10.1182/blood.V96.12.3725.h8003725_3725_3733}},
volume = {{96}},
year = {{2000}},
}