“Background”. Human Herpes virus 6 (HHV-6), a lymphotropic virus, infects and establishes a latent infection in endothelial cells (ECs). Following HHV-6 infection HHV-6 infection induces the loss of angiogenic properties in ECs as shown by inability to form capillary-like structures and seal wound scratches. The antiangiogenic effects observed in infected ECs are associated to the expression of HHV-6 U94/rep, a latency-associated gene. “Methods”. The full length U94 gene was cloned in the pVAX plasmid. Human umbilical ECs (HUVECs) were isolated, characterized, and cultured. HUVECs were transfected with an endotoxin-free U94-expressing pVAX plasmid by AMAXA. An empty U94 vector was used as control in all experiments. “Results”. Here we show that HUVECs expressing U94 are inhibited in their migratory capacity as demonstrated by “wound sealing” assay. Differently from cells nucleofected with empty pVAX, HUVECs expressing U94 were completely unable to repair a wound scratch carried out on a confluent cellular monolayer. Moreover, cells expressing U94 loss their ability to form a network of capillary-like structures when seeded on extracellular matrix (BME) coated plates. “Conclusions”. The capability of HHV-6 U94 to block both migratory and angiogenic capability of ECs may lead, if confirmed by experimental animal models, to the development of new therapeutic approaches in all those diseases in which an aberrant angiogenesis need to be modulated.
U94 gene of HHV-6 inhibits migration and angiogenesis in human endothelial cells
CASELLI, Elisabetta;DI LUCA, Dario;
2011
Abstract
“Background”. Human Herpes virus 6 (HHV-6), a lymphotropic virus, infects and establishes a latent infection in endothelial cells (ECs). Following HHV-6 infection HHV-6 infection induces the loss of angiogenic properties in ECs as shown by inability to form capillary-like structures and seal wound scratches. The antiangiogenic effects observed in infected ECs are associated to the expression of HHV-6 U94/rep, a latency-associated gene. “Methods”. The full length U94 gene was cloned in the pVAX plasmid. Human umbilical ECs (HUVECs) were isolated, characterized, and cultured. HUVECs were transfected with an endotoxin-free U94-expressing pVAX plasmid by AMAXA. An empty U94 vector was used as control in all experiments. “Results”. Here we show that HUVECs expressing U94 are inhibited in their migratory capacity as demonstrated by “wound sealing” assay. Differently from cells nucleofected with empty pVAX, HUVECs expressing U94 were completely unable to repair a wound scratch carried out on a confluent cellular monolayer. Moreover, cells expressing U94 loss their ability to form a network of capillary-like structures when seeded on extracellular matrix (BME) coated plates. “Conclusions”. The capability of HHV-6 U94 to block both migratory and angiogenic capability of ECs may lead, if confirmed by experimental animal models, to the development of new therapeutic approaches in all those diseases in which an aberrant angiogenesis need to be modulated.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
