Retroviral display of urokinase-binding domain fused to amphotropic envelope protein.
Biochem Biophys Res Commun
; 331(4): 1485-93, 2005 Jun 17.
Article
en En
| MEDLINE
| ID: mdl-15883041
Tumors frequently express urokinase (uPA) receptor (uPAR). To investigate whether uPAR can efficiently target cancerous cells using amphotropic retroviral vectors, we generated a retrovirus displaying the amino-terminal fragment (ATF) of uPA as an N-terminal extension of viral envelope protein. We also made use of a "two-step strategy" by inserting a uPA cleavage site between the ATF moiety and the envelope. We measured the ability of ATF-bearing chimeric envelopes to infect huPAR-overexpressing Madin-Darby canine kidney (MDCK) and control MDCK II cells. The ATF-viruses infected both MDCK cell lines with an equivalent efficiency, suggesting that the chimeric viruses were not sequestered by uPAR and infect cells preferentially via the Pit-2 receptor. The addition of a uPA cleavage site increased the infection level of huPAR-MDCK cells by 2-fold when uPA was present in the infection medium. Surprisingly, ATF-env viruses infected huPAR-MDCK cells 5.5-fold more efficiently in the presence of exogenous uPA. This stimulatory effect of uPA on infection of huPAR-MDCK cells by the ATF-env virus was completely abolished by methyl-beta-cyclodextrin, suggesting that this effect involves the caveolar endocytosis pathway.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Retroviridae
/
Activador de Plasminógeno de Tipo Uroquinasa
/
Proteínas del Envoltorio Viral
Límite:
Animals
Idioma:
En
Revista:
Biochem Biophys Res Commun
Año:
2005
Tipo del documento:
Article
País de afiliación:
Francia
Pais de publicación:
Estados Unidos