In vivo image analysis of BoHV-4-based vector in mice

• Published in: PloS one 2014-04, Vol.9 (4), p.e95779-e95779
• Main Authors: Franceschi, Valentina, Stellari, Fabio Franco, Mangia, Carlo, Jacca, Sarah, Lavrentiadou, Sophia, Cavirani, Sandro, Heikenwalder, Mathias, Donofrio, Gaetano
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis; Artificial chromosomes; Bacteria; Bacterial artificial chromosomes; Biology and Life Sciences; Bison bison; Buffalo; Cattle; Cell culture; Cell Line; Cloning; Cytomegalovirus; Dogs; Drug delivery systems; Enzymes; Female; Gene therapy; Gene transfer; Gene Transfer Techniques; Genetic aspects; Genetic vectors; Genetic Vectors - genetics; Genomes; Hepatocytes; Herpesvirus 4, Bovine - genetics; Herpesviruses; Histochemistry; Humans; Image analysis; Image detection; Image processing; Immunohistochemistry; In vivo methods and tests; Inoculation; Intravenous administration; Kinases; Liver; Mice; Physiological aspects; Replication; Research and Analysis Methods; Science; Sheep; Viruses
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0095779

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.