Retroviral Vector-Mediated Transfer and Expression of Human Tissue Plasminogen Activator cDNA in Bovine Brain Endothelial Cells
作者:
Hong Yu,
Ying Wang,
Darwin Eton,
Monique Stins,
Liang Wang,
Michael Apuzzo,
Fred Weaver,
J. McComb,
Martin Weiss,
Berislav Zlokovic,
&NA; &NA;,
期刊:
Neurosurgery
(OVID Available online 1999)
卷期:
Volume 45,
issue 4
页码: 962-962
ISSN:0148-396X
年代: 1999
出版商: OVID
关键词: Brain endothelial cells;Gene transfer;Plasminogen activator;Retroviral vector
数据来源: OVID
摘要:
OBJECTIVESGene transfer of thrombolytic enzymes to vascular endothelial cells may influence the kinetics of intravascular thrombosis. This study defines the potential for gene transfer of tissue plasminogen activator (tPA) into bovine brain endothelial cells (BBEC).METHODSThe retroviral vectors derived from murine leukemia virus (MuLV) were used to transfer human tPA cDNA to BBEC. The tPA activity, tPA antigen and tPA inhibitor 1 (PAI-1) antigen were determined in the supernatant of transduced (BBEC/tPA) cell cultures by an immunoassay.RESULTSThe tPA antigen and enzymatic activity in cell culture supernatants of BBEC/tPA transduced cells were 75 ng/ml and 14 IU/ml after 4 days, that was 25 and 28-fold higher compared to the respective values in control cells. The PAI-1 antigen was not affected by tPA cDNA transfer. The Western blot assay of cell lysates confirmed that the majority of tPA in BBEC/tPA transduced cells was in the form of free tPA. While the maximal transduction efficiency of BBEC with an amphotropic MuLV vector was about 15%, a MuLV pseudotyped with vesicular stomatitis virus G glycoprotein envelope achieved high > 90% maximal transduction efficiency.CONCLUSIONSThe fibrinolytic activity of brain endothelial cells can be enhanced by transferring human tPA cDNA. These findings provide an initial step in implementation of future studies that investigate the use of this technology as an adjunctive treatment for cerebrovascular disease.
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