Transduction of the rat brain by Bovine Herpesvirus 4
© Redaelli et al; licensee BioMed Central Ltd. 2008
Received: 30 October 2007
Accepted: 12 February 2008
Published: 12 February 2008
Bovine herpesvirus 4 (BoHV-4) is a gamma-herpesvirus with no clear disease association. A recombinant BoHV-4 (BoHV-4EGFPΔTK) expressing Green Fluorescent Protein (EGFP), was successfully used to infect F98 rat glioma cells. BoHV-4EGFPΔTK was injected into the lateral ventricle of the rat brain. Histology and immunohistochemistry showed that ependymal and rostral migratory stream cells were transduced while neurons were not. Clinical scores, evaluated for 90 days, indicated that the virus was non neuropathogenic, suggesting this virus is a suitable vector for brain tumor gene therapy.
Gene delivery and targeting is a major issue in the treatment of severe brain tumors. The cancer treatment mediated or coadiuvated by genetically modified oncolytic viruses is an interesting opportunity in clinical oncology.
Bovine herpesvirus 4 (BoHV-4) is a gamma-herpesvirus with no clear disease association , suggesting it as a suitable vector for gene therapy. BoHV-4 has been isolated from different tissues and has been show to establish a persistent infection in its natural host, the cattle, and in the experimental animal, the rabbit [2, 3]. In the natural and experimental host some evidence indicates that the monocyte/macrophage lineage is a site of persistent infection [4, 5]. Interestingly, unlike other gamma-herpes viruses like Epstein-Barr Virus  and Herpes Virus Saimiri , BoHV-4 is non oncogenic. Hence, BoHV-4 could be employed as a possible therapeutic candidate as attenuation of genes to render it non-pathogenic is not required. However, BoHV-4 does replicate and cause a cytopathic effect in a number of immortalized cell lines and primary cell cultures [8, 9].
We have previously demonstrated that BoHV-4 does not replicate in the mouse brain and that infection was restricted to ependymal and rostral migratory stream (RMS) regions after viral injection in the lateral ventricle of the mouse brain . The aim of this work was to evaluate the suitability of BoHV-4 as a vector for glioma gene therapy. The virus was first assessed in vitro, using the rat glioma F98 cell line (ATCC, USA) and then in vivo by injecting the virus into the brain of rats.
In vitro, BoHV-4 is able to replicate in primary cell culture or cell lines from a broad spectrum of host species. The infection in some permissive cells leads to viral progeny and CPE; in other cells, although CPE takes place, no viral progeny is produced; whereas in some non permissive cells BoHV-4 infection is persistent with no effect on cell survival . The nature of cell death induced by BoHV-4 is highly controversial. For some cell types, it is mediated by apoptosis [12–14], but in other cells BoHV-4 infection protects against TNF-alpha induced apoptosis . Since BoHV-4 induced CPE in F98 cells, the nature of cell death was investigated. F98 cells were infected with 5 m.o.i. of BoHV-4 and cell death was examined by Wright's nuclear staining with propidium iodide and by internucleosomal DNA fragmentation. Both approaches showed that BoHV-4-induced CPE was not mediated by apoptosis (data not shown).
The most interesting observation made during this study, was the ability of BoHV-4EGFPΔTK to replicate in highly replicating glioma cells but not in post mitotic brain cells. This observation could be explained by a proteomic switch occurring in the intracellular microenvironment of tumor cells capable of activating the full replication cycle of BoHV-4EGFPΔTK.
The absence of pathogenicity in the rat brain and the ability to establish a permissive infection in cultures of glioma cells, make BoHV-4 an ideal candidate as a gene delivery or oncolytic vector for gliomas in the nervous system.
We would like to thank Dr. Shan Herath (university of London) for English editing of the manuscript and Italian Minister of Science (Prin 2005) and Fondazione CARIPARMA for financial support.
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