Bovine herpesvirus 4 based vector interaction with liver cells in vitro and in vivo.
Gene transfer into hepatocytes is highly desirable for the long-term goal of replacing deficient proteins and correcting metabolic disorders. Bovine herpesvirus 4 (BoHV-4) based vector capability to transduce rat liver cells in vitro and in vivo was assessed. For the in vitro study, a buffalo rat liver cell line was successfully transduced by BoHV-4 and although did not show toxicity, the immediate early two viral gene was transcribed and cells harboring the intact viral genome could be pharmacologically selected, but no viral replication took place. For the in vivo study, adult male rats were inoculated intraportally and intraparenchimally with a BoHV-4 expressing enhanced green fluorescent protein and liver sections were analyzed through fluorescent microscopy. Although the liver parenchyma could not be transduced, the endothelial layer of the liver vasculature showed a robust transgene expression without toxicity. Successful BoHV-4 based vector transduction of primary cultures of rat hepatocytes suggests that extrinsic factors, and not hepatocytes per se, are the cause of such lack of transducibility. The present study serves as a starting point for study of the use of BoHV-4 based vectors to target gene delivery to vascular endothelial cells.
Donofrio
Gaetano
G
Dipartimento di Salute Animale, Facoltà di Medicina Veterinaria, Sezione di Malattie Infettive degli Animali, Università di Parma, via del Taglio 8, 43100 Parma, Italy. gaetano.donofrio@unipr.it
Martignani
Eugenio
E
Poli
Enzo
E
Lange
Claudia
C
Martini
Filippo Maria
FM
Cavirani
Sandro
S
Cabassi
Clotilde Silvia
CS
Taddei
Simone
S
Flammini
Cesidio Filippo
CF
D000818:Animals; D002460:Cell Line; D042783:Endothelial Cells / Q000821:virology; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000009:adverse effects; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D022781:Hepatocytes / Q000821:virology*; D026021:Herpesvirus 4, Bovine / Q000235:genetics* / Q000254:growth & development; D008099:Liver / Q000821:virology; D008297:Male; D008856:Microscopy, Fluorescence; D051381:Rats; D011915:Rats, Inbred BUF; D013194:Staining and Labeling / Q000379:methods
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins