Abstract
Human immunodeficiency virus (HIV, lentivirus) vector has attractive features for gene therapy, including the ability to transduce non-dividing cells and long-term transgene expression. We have already reported that lentivirus vector can transduce well-differentiated rat cardiac myocytes. Endothelial cells (EC) are an attractive target for gene therapy, both for the treatment of cardiovascular disease and for the systemic delivery of recombinant gene products directly into the circulation. There are several reports regarding application of adenovirus and retrovirus based vectors to EC. However, there have been few reports which show the effect to lentivirus-mediated gene transfer efficiency, compared with adenovirus and retrovirus. In this study, bovine aortic endothelial cells (BAECs) were infected, in vitro, with these virus vectors. Transduction efficiency (TE) of β-Gal gene transfer in BAECs by adenovirus, lentivirus, or retrovirus at MOI10 (Multiplicity of infection) (determined on Hela cells) is 69±11, 33±8, or 22±6% respectively. In adenovirus and lentivirus, almost 100% of BAECs were transduced at MOI 50. However, in retrovirus, TE showed only 48±6% at MOI 50 and no increase at MOI 100. The percentage of β-Gal positive cells was decreased rapidly at longer passage of cells after being transduced by adenovirus. However, lentivirus and retrovirus showed sustained higher percentage of positive cells. Furthermore, transduction by lentiviral vectors had no significant effect on viability of BAECs. Our results indicate that lentivirus showed high-level and long term gene expression in BAECs. Lentivirus can be an effective vector for the ex vivo, genetically modified EC implantation and in vivo gene therapy.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1-11 |
| Number of pages | 11 |
| Journal | Preparative Biochemistry and Biotechnology |
| Volume | 37 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jan 18 2007 |
| Externally published | Yes |
Fingerprint
Keywords
- Adenovirus
- Endotherial cells
- Gene therapy
- Lentivirus
- Retrovirus
ASJC Scopus subject areas
- Biochemistry
- Biotechnology
- Molecular Biology
- Applied Microbiology and Biotechnology
Cite this
Lentiviral vector-mediated gene transfer to endotherial cells compared with adenoviral and retroviral vectors. / Sakoda, Tsuyoshi; Kasahara, Noriyuki; Kedes, Larry; Ohyanagi, Mitsumasa.
In: Preparative Biochemistry and Biotechnology, Vol. 37, No. 1, 18.01.2007, p. 1-11.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Lentiviral vector-mediated gene transfer to endotherial cells compared with adenoviral and retroviral vectors
AU - Sakoda, Tsuyoshi
AU - Kasahara, Noriyuki
AU - Kedes, Larry
AU - Ohyanagi, Mitsumasa
PY - 2007/1/18
Y1 - 2007/1/18
N2 - Human immunodeficiency virus (HIV, lentivirus) vector has attractive features for gene therapy, including the ability to transduce non-dividing cells and long-term transgene expression. We have already reported that lentivirus vector can transduce well-differentiated rat cardiac myocytes. Endothelial cells (EC) are an attractive target for gene therapy, both for the treatment of cardiovascular disease and for the systemic delivery of recombinant gene products directly into the circulation. There are several reports regarding application of adenovirus and retrovirus based vectors to EC. However, there have been few reports which show the effect to lentivirus-mediated gene transfer efficiency, compared with adenovirus and retrovirus. In this study, bovine aortic endothelial cells (BAECs) were infected, in vitro, with these virus vectors. Transduction efficiency (TE) of β-Gal gene transfer in BAECs by adenovirus, lentivirus, or retrovirus at MOI10 (Multiplicity of infection) (determined on Hela cells) is 69±11, 33±8, or 22±6% respectively. In adenovirus and lentivirus, almost 100% of BAECs were transduced at MOI 50. However, in retrovirus, TE showed only 48±6% at MOI 50 and no increase at MOI 100. The percentage of β-Gal positive cells was decreased rapidly at longer passage of cells after being transduced by adenovirus. However, lentivirus and retrovirus showed sustained higher percentage of positive cells. Furthermore, transduction by lentiviral vectors had no significant effect on viability of BAECs. Our results indicate that lentivirus showed high-level and long term gene expression in BAECs. Lentivirus can be an effective vector for the ex vivo, genetically modified EC implantation and in vivo gene therapy.
AB - Human immunodeficiency virus (HIV, lentivirus) vector has attractive features for gene therapy, including the ability to transduce non-dividing cells and long-term transgene expression. We have already reported that lentivirus vector can transduce well-differentiated rat cardiac myocytes. Endothelial cells (EC) are an attractive target for gene therapy, both for the treatment of cardiovascular disease and for the systemic delivery of recombinant gene products directly into the circulation. There are several reports regarding application of adenovirus and retrovirus based vectors to EC. However, there have been few reports which show the effect to lentivirus-mediated gene transfer efficiency, compared with adenovirus and retrovirus. In this study, bovine aortic endothelial cells (BAECs) were infected, in vitro, with these virus vectors. Transduction efficiency (TE) of β-Gal gene transfer in BAECs by adenovirus, lentivirus, or retrovirus at MOI10 (Multiplicity of infection) (determined on Hela cells) is 69±11, 33±8, or 22±6% respectively. In adenovirus and lentivirus, almost 100% of BAECs were transduced at MOI 50. However, in retrovirus, TE showed only 48±6% at MOI 50 and no increase at MOI 100. The percentage of β-Gal positive cells was decreased rapidly at longer passage of cells after being transduced by adenovirus. However, lentivirus and retrovirus showed sustained higher percentage of positive cells. Furthermore, transduction by lentiviral vectors had no significant effect on viability of BAECs. Our results indicate that lentivirus showed high-level and long term gene expression in BAECs. Lentivirus can be an effective vector for the ex vivo, genetically modified EC implantation and in vivo gene therapy.
KW - Adenovirus
KW - Endotherial cells
KW - Gene therapy
KW - Lentivirus
KW - Retrovirus
UR - http://www.scopus.com/inward/record.url?scp=33845437506&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33845437506&partnerID=8YFLogxK
U2 - 10.1080/10826060601039345
DO - 10.1080/10826060601039345
M3 - Article
C2 - 17134978
AN - SCOPUS:33845437506
VL - 37
SP - 1
EP - 11
JO - Preparative Biochemistry and Biotechnology
JF - Preparative Biochemistry and Biotechnology
SN - 1082-6068
IS - 1
ER -