Abstract
Due to its biological characteristics bovine herpesvirus 4 (BoHV-4) has been considered as an appropriate gene delivery vector. Its genomic clone, modified as a bacterial artificial chromosome (BAC), is better genetically manipulable and can be used as an efficient gene delivery and vaccine vector. Although a large amount of data have been accumulated in vitro on this specific aspect, the same cannot be asserted for the in vivo condition. Therefore, here we investigated the fate of a recombinant BoHV-4 strain expressing luciferase (BoHV-4-A-CMVlucΔTK) after intraperitoneal or intravenous inoculation in mice, by generating a novel recombinant BoHV-4 expressing luciferase (BoHV-4-A-CMVlucΔTK) and by following the virus replication through in vivo imaging analysis. BoHV-4-A-CMVlucΔTK was first characterized in vitro where it was shown, on one hand that its replication properties are identical to those of the parental virus, and on the other that the transduced/infected cells strongly express luciferase. When BoHV-4-A-CMVlucΔTK was inoculated in mice, either intraperitoneally or intravenously, BoHV-4-A-CMVlucΔTK infection/transduction was exclusively localized to the liver, as detected by in vivo image analysis, and in particular almost exclusively in the hepatocytes, as determined by immuno-histochemistry. These data, that add a new insight on the biology of BoHV-4 in vivo, provide the first indication for the potential use of a BoHV-4-based vector in gene-transfer in the liver.
Highlights
Bovine herpesvirus 4 (BoHV-4) is a gammaherpesvirus able to replicate in a broad range of host species both in vivo and in vitro [1]
In addition to its natural hosts, BoHV-4 establishes persistent infections in the rabbit, which is considered as an experimental host of the virus [8,9] The BoHV-4-based vector, a recombinant BoHV-4 cloned as bacterial artificial chromosome (BAC) expressing diverse immune-dominant antigens from different pathogens, has been shown to successfully immunize mice [10], rats [11], rabbits [5], sheep [3], swine [6] and goats [12]
To overcome to some extent this gap, in the present study we investigated by novel in vivo image analysis techniques, transduction and clearances of BoHV-4 in mice experimentally infected with a novel recombinant BoHV-4 (BoHV-4-A-CMVlucDTK) expressing firefly luciferase
Summary
Bovine herpesvirus 4 (BoHV-4) is a gammaherpesvirus able to replicate in a broad range of host species both in vivo and in vitro [1]. BoHV-4 is able to replicate in primary cell cultures and cell lines from various animal species such as cattle, sheep, goat, swine, cat, dog, rabbit, mink, horse, turkey, ferret, chicken, hamster, rat, mouse, and monkey [1,2,3,4,5,6,7]. In addition to its natural hosts, BoHV-4 establishes persistent infections in the rabbit, which is considered as an experimental host of the virus [8,9] The BoHV-4-based vector, a recombinant BoHV-4 cloned as bacterial artificial chromosome (BAC) expressing diverse immune-dominant antigens from different pathogens, has been shown to successfully immunize mice [10], rats [11], rabbits [5], sheep [3], swine [6] and goats [12]. To overcome to some extent this gap, in the present study we investigated by novel in vivo image analysis techniques, transduction and clearances of BoHV-4 in mice experimentally infected with a novel recombinant BoHV-4 (BoHV-4-A-CMVlucDTK) expressing firefly luciferase
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