This Article Full Text (PDF) References Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Add to Saved Citations Download to citation manager Request Permissions Request Reprints Add to My Marked Citations Citing Articles Citing Articles via Google Scholar Citing Articles via Scopus Google Scholar Articles by Ai Kim, J. Articles by Fisher, M. J. Search for Related Content PubMed PubMed Citation Articles by Ai Kim, J. Articles by Fisher, M. J. Social Bookmarking                   What's this?

Adenoviral-Mediated Transfer of Tissue Plasminogen Activator Gene into Brain Capillary Endothelial Cells In Vitro

Catherine C. Hedrick, PhD

Dangci Xie, PhD

Mark J. Fisher, MD

Tissue plasminogen activator (tPA) has a critical role in fibrinolysis, converting plasminogen into active protease plasmin. Because intravenous tPA has only limited effectiveness as acute stroke therapy, enhancement of endogenous tPA represents a potential alternative to stroke treatment. Adenoviral-mediated gene transfer was used to enhance production of tPA in bovine brain capillary endothelial cells (BEC). Antigen and activity levels of tPA and plasminogen activator inhibitor-1 (PAI-1) in media from BEC infected with AdCMVtPA were analyzed. Conditioned media were analyzed for thrombomodulin, the integral membrane antithrombotic molecule that co-activates protein C. BEC infected with AdCMVtPA demonstrated enhanced expression of tPA antigen (40.2 ±0.4 ng/mL vs 1.1 ±1.5 ng/mL [p<0.001] and 0.3 ±0.5 ng/mL [p<0.0001 ], respectively) and increased tPA enzymatic activity (27.4 ±5.7 IU/mL vs 8.3 ±1.7 IU/mL [p<0.05] and 13.3 ±3.2 IU/mL [p<0.05], respectively) compared to BEC infected with the control adenovirus (AdI327) or uninfected BEC. There was a moderate increase in PAI-1 protein 4 days after transfection with AdCMVtPA, and the integral membrane protein throm bomodulin was released into media by transfected BEC. These results demonstrate that aden oviral-mediated delivery in vitro of the human tPA gene resulted in high levels of expression of tPA in BEC. Transient overexpression of tPA by gene transfer might be a useful strategy to protect against thrombotic occlusion during the period of risk of acute stroke.

CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?