title: In vivo attenuation of recombinant murine gammaherpesvirus 68 (MHV-68) is due to the expression and immunogenicity but not to the insertion of foreign sequences.
abstract: Recombinant herpesviruses are increasingly utilized to study herpesvirus biology. For recombinant viruses carrying insertions of foreign sequences, attenuated phenotypes in vivo have been frequently observed. In most cases, the underlying mechanisms were not clear or have not been investigated. In this study, we used a recombinant murine gammaherpesvirus 68 (MHV-68), carrying a cassette for the expression of the non-structural protein NS3 of Hepatitis C virus (MHV-68-NS3), to systematically address the question whether the insertion of a defined foreign sequence (NS3) interferes with the biological properties of the recombinant virus in vivo, and to analyze the underlying mechanism. We show that while MHV-68-NS3 is attenuated in vivo, recombinant MHV-68 carrying identical genomic inserts but unable to express the NS3 protein, are not attenuated. Moreover, we provide evidence that the attenuated phenotype of MHV-68-NS3 is caused by the immune response. Our findings are important for the in vivo use of recombinant MHV-68 carrying insertions of marker genes, reporter genes or genes of model antigens. They are also relevant for the potential application of MHV-68 as gene delivery vector.
authors.affiliation: Institute for Virology, Technical University, München, Germany.
authors.forename: Susanne
authors.initials: S
authors.lastname: El-Gogo
authors.forename: Britta
authors.initials: B
authors.lastname: Flach
authors.forename: Caroline
authors.initials: C
authors.lastname: Staib
authors.forename: Gerd
authors.initials: G
authors.lastname: Sutter
authors.forename: Heiko
authors.initials: H
authors.lastname: Adler
mesh_terms: D000818:Animals; D005260:Female; D015870:Gene Expression; D018937:Hepatitis C Antibodies / Q000276:immunology*; D008168:Lung / Q000821:virology; D051379:Mice; D008810:Mice, Inbred C57BL; D016254:Mutagenesis, Insertional; D011995:Recombination, Genetic; D018144:Rhadinovirus / Q000235:genetics / Q000276:immunology* / Q000472:pathogenicity*; D013154:Spleen / Q000473:pathology; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D018937:Hepatitis C Antibodies; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases