|
|
 Previous Article | Table of Contents | Next Article 
Enhancement of the fibrinolytic activity of sheep endothelial cells by
retroviral vector-mediated gene transfer
DA Dichek, O Nussbaum, SJ Degen and WF Anderson
Molecular Hematology Branch, National Heart, Lung and Blood Institute,
National Institutes of Health, Bethesda, MD 20892.
In an attempt to enhance the fibrinolytic activity of endothelial cells
(EC), a retroviral vector containing the human tissue-type plasminogen
activator (t-PA) cDNA was constructed. Sheep EC were stably transduced with
the vector, resulting in a 30-fold increase in t-PA activity over that
detected in EC transduced with a control vector. Southern and Northern
analyses confirmed the presence of both the vector sequence and the
appropriate mRNA transcripts. Secretion of high levels of recombinant human
t-PA continued in vitro for the duration of the experiments, up to 11 weeks
after transduction, although the rate of t- PA secretion decreased in some
of the EC lines. Zymographic analysis of conditioned medium from
t-PA-transduced EC showed the presence of two new molecular species with
plasminogen activator activity that could be specifically
immunoprecipitated with a monoclonal antihuman t-PA antibody. The relative
molecular masses of these species (60 to 80 and 110 Kd) suggest that they
represent recombinant human t-PA both free and bound to sheep plasminogen
activator inhibitor 1 (PAI-1). Consistent with this interpretation, the
110-Kd species could be specifically immunoprecipitated with antiserum to
PAI-1. These studies demonstrate that retroviral vector-mediated gene
transfer may be used to increase total EC fibrinolytic activity.
Volume 77,
Issue 3,
pp. 533-541,
02/01/1991
Copyright © 1991 by The American Society of Hematology
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
Z. S. Katusic, N. M. Caplice, and K. A. Nath
Nitric Oxide Synthase Gene Transfer as a Tool to Study Biology of Endothelial Cells
Arterioscler. Thromb. Vasc. Biol.,
November 1, 2003;
23(11):
1990 - 1994.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Ai Kim, C. C. Hedrick, Dangci Xie, and M. J. Fisher
Adenoviral-Mediated Transfer of Tissue Plasminogen Activator Gene into Brain Capillary Endothelial Cells In Vitro
Angiology,
September 1, 2001;
52(9):
627 - 634.
[Abstract]
[PDF]
|
|
|
|
|
|
|
I. J. Kullo, R. D. Simari, and R. S. Schwartz
Vascular Gene Transfer : From Bench to Bedside
Arterioscler. Thromb. Vasc. Biol.,
February 1, 1999;
19(2):
196 - 207.
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. F. Dunn, K. D. Newman, M. Jones, I. Yamada, V. Shayani, R. Virmani, and D. A. Dichek
Seeding of Vascular Grafts With Genetically Modified Endothelial Cells : Secretion of Recombinant TPA Results in Decreased Seeded Cell Retention In Vitro and In Vivo
Circulation,
April 1, 1996;
93(7):
1439 - 1446.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
D. A. Dichek, J. Anderson, A. B. Kelly, S. R. Hanson, and L. A. Harker
Enhanced In Vivo Antithrombotic Effects of Endothelial Cells Expressing Recombinant Plasminogen Activators Transduced With Retroviral Vectors
Circulation,
January 15, 1996;
93(2):
301 - 309.
[Abstract]
[Full Text]
|
|
|
|
|
