Retrovirus-mediated transfer of a hygromycin phosphotransferase-thymidine kinase fusion gene into human CD34 + bone marrow cells

Yoshiki Akatsuka, Nobuhiko Emi, Hidefumi Kato, Akihiro Abe, Mitsune Tanimoto, Stephen D. Lupton, Hidehiko Saito

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Retrovirus-mediated gene transfer into human hematopoietic stem cells has been proposed as a means of therapy for various inherited diseases and as a method of gene marking. The transduction efficiency of an amphotropic retroviral vector (PA317/HyTK) containing a hygromycin phosphotransferase-thymidine kinase fusion gene was examined with human CD34 + bone marrow cells in the presence of interleukin-3 (IL-3), interleukin-6 (IL-6), and stem cell factor. Transduction efficiencies determined from the ability of transduced granulocyte-macrophage colony forming units (CFU-GM) to grow in hygromycin B and from polymerase chain reaction analysis of individual transduced CFU-GM growing in the presence of hygromycin B were 0.3-3.0% (mean ± S.D., 1.1 ± 0.9%) and 0.1-1.2% (mean ± S.D., 0.5 ± 0.4%), respectively. Ganciclovir at a dose of ∼ 1 μM reduced the number of CFU-GM derived from vector-infected CD34 + cells by 50%. These findings demonstrate that human hematopoietic stem cells infected with this retroviral vector are susceptible to ganciclovir, offering the potential to control transduced gene expression in vivo.

Original languageEnglish
Pages (from-to)251-261
Number of pages11
JournalInternational Journal of Hematology
Volume60
Issue number4
DOIs
Publication statusPublished - 01-12-1994
Externally publishedYes

Fingerprint

hygromycin-B kinase
Granulocyte-Macrophage Progenitor Cells
Thymidine Kinase
Gene Fusion
Retroviridae
Bone Marrow Cells
Hygromycin B
Ganciclovir
Hematopoietic Stem Cells
Stem Cell Factor
Interleukin-3
Genes
Interleukin-6
Gene Expression
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Hematology

Cite this

Akatsuka, Yoshiki ; Emi, Nobuhiko ; Kato, Hidefumi ; Abe, Akihiro ; Tanimoto, Mitsune ; Lupton, Stephen D. ; Saito, Hidehiko. / Retrovirus-mediated transfer of a hygromycin phosphotransferase-thymidine kinase fusion gene into human CD34 + bone marrow cells In: International Journal of Hematology. 1994 ; Vol. 60, No. 4. pp. 251-261.
@article{0654b985985e48ca812f5e0139eb8c66,
title = "Retrovirus-mediated transfer of a hygromycin phosphotransferase-thymidine kinase fusion gene into human CD34 + bone marrow cells",
abstract = "Retrovirus-mediated gene transfer into human hematopoietic stem cells has been proposed as a means of therapy for various inherited diseases and as a method of gene marking. The transduction efficiency of an amphotropic retroviral vector (PA317/HyTK) containing a hygromycin phosphotransferase-thymidine kinase fusion gene was examined with human CD34 + bone marrow cells in the presence of interleukin-3 (IL-3), interleukin-6 (IL-6), and stem cell factor. Transduction efficiencies determined from the ability of transduced granulocyte-macrophage colony forming units (CFU-GM) to grow in hygromycin B and from polymerase chain reaction analysis of individual transduced CFU-GM growing in the presence of hygromycin B were 0.3-3.0{\%} (mean ± S.D., 1.1 ± 0.9{\%}) and 0.1-1.2{\%} (mean ± S.D., 0.5 ± 0.4{\%}), respectively. Ganciclovir at a dose of ∼ 1 μM reduced the number of CFU-GM derived from vector-infected CD34 + cells by 50{\%}. These findings demonstrate that human hematopoietic stem cells infected with this retroviral vector are susceptible to ganciclovir, offering the potential to control transduced gene expression in vivo.",
author = "Yoshiki Akatsuka and Nobuhiko Emi and Hidefumi Kato and Akihiro Abe and Mitsune Tanimoto and Lupton, {Stephen D.} and Hidehiko Saito",
year = "1994",
month = "12",
day = "1",
doi = "10.1016/0925-5710(94)00334-B",
language = "English",
volume = "60",
pages = "251--261",
journal = "International Journal of Hematology",
issn = "0925-5710",
publisher = "Springer Japan",
number = "4",

}

Retrovirus-mediated transfer of a hygromycin phosphotransferase-thymidine kinase fusion gene into human CD34 + bone marrow cells . / Akatsuka, Yoshiki; Emi, Nobuhiko; Kato, Hidefumi; Abe, Akihiro; Tanimoto, Mitsune; Lupton, Stephen D.; Saito, Hidehiko.

In: International Journal of Hematology, Vol. 60, No. 4, 01.12.1994, p. 251-261.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Retrovirus-mediated transfer of a hygromycin phosphotransferase-thymidine kinase fusion gene into human CD34 + bone marrow cells

AU - Akatsuka, Yoshiki

AU - Emi, Nobuhiko

AU - Kato, Hidefumi

AU - Abe, Akihiro

AU - Tanimoto, Mitsune

AU - Lupton, Stephen D.

AU - Saito, Hidehiko

PY - 1994/12/1

Y1 - 1994/12/1

N2 - Retrovirus-mediated gene transfer into human hematopoietic stem cells has been proposed as a means of therapy for various inherited diseases and as a method of gene marking. The transduction efficiency of an amphotropic retroviral vector (PA317/HyTK) containing a hygromycin phosphotransferase-thymidine kinase fusion gene was examined with human CD34 + bone marrow cells in the presence of interleukin-3 (IL-3), interleukin-6 (IL-6), and stem cell factor. Transduction efficiencies determined from the ability of transduced granulocyte-macrophage colony forming units (CFU-GM) to grow in hygromycin B and from polymerase chain reaction analysis of individual transduced CFU-GM growing in the presence of hygromycin B were 0.3-3.0% (mean ± S.D., 1.1 ± 0.9%) and 0.1-1.2% (mean ± S.D., 0.5 ± 0.4%), respectively. Ganciclovir at a dose of ∼ 1 μM reduced the number of CFU-GM derived from vector-infected CD34 + cells by 50%. These findings demonstrate that human hematopoietic stem cells infected with this retroviral vector are susceptible to ganciclovir, offering the potential to control transduced gene expression in vivo.

AB - Retrovirus-mediated gene transfer into human hematopoietic stem cells has been proposed as a means of therapy for various inherited diseases and as a method of gene marking. The transduction efficiency of an amphotropic retroviral vector (PA317/HyTK) containing a hygromycin phosphotransferase-thymidine kinase fusion gene was examined with human CD34 + bone marrow cells in the presence of interleukin-3 (IL-3), interleukin-6 (IL-6), and stem cell factor. Transduction efficiencies determined from the ability of transduced granulocyte-macrophage colony forming units (CFU-GM) to grow in hygromycin B and from polymerase chain reaction analysis of individual transduced CFU-GM growing in the presence of hygromycin B were 0.3-3.0% (mean ± S.D., 1.1 ± 0.9%) and 0.1-1.2% (mean ± S.D., 0.5 ± 0.4%), respectively. Ganciclovir at a dose of ∼ 1 μM reduced the number of CFU-GM derived from vector-infected CD34 + cells by 50%. These findings demonstrate that human hematopoietic stem cells infected with this retroviral vector are susceptible to ganciclovir, offering the potential to control transduced gene expression in vivo.

UR - http://www.scopus.com/inward/record.url?scp=0028607646&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028607646&partnerID=8YFLogxK

U2 - 10.1016/0925-5710(94)00334-B

DO - 10.1016/0925-5710(94)00334-B

M3 - Article

VL - 60

SP - 251

EP - 261

JO - International Journal of Hematology

JF - International Journal of Hematology

SN - 0925-5710

IS - 4

ER -