title: Interferon-containing supernatants increase Marek's disease herpesvirus genomes and gene transcription levels, but not virion replication in vitro.
abstract: Viruses encounter the innate immune system immediately after infection of the host; specifically, soluble molecules that are both directly lethal and that initiate acquired immunity. Using the oncogenic Marek's disease alpha-herpesvirus (MDV) model, we quantified the effect of a interferon-containing supernatants (ICS), on MDV replication, gene transcription and antigen expression kinetics. We used an established cell culture system and a well-defined virulent MDV (RB-1B). RB-1B was cultured without ICS, or pretreated and then continuously treated with ICS. We compared (i) RB-1B infectivity; (ii) RB-1B growth by microscopy; (iii) numbers of cells expressing RB-1B antigens by flow cytometry; (iv) RB-1B-DNA load per cell by duplex real-time PCR, and (v) gene transcription kinetics for key MDV-life stages by duplex real-time reverse-transcriptase PCR (RT-PCR). ICS inhibited RB-1B infection, completion of productive life cycle and cell-to-cell infection. The numbers of cells expressing glycoprotein B (a kinetically late antigen) greatly decreased, but the numbers of cells expressing pp38 (a kinetically early antigen) decreased only slightly. The two greatest effects were increases in both RB-1B-DNA per infected cell and pp38 mRNA. We propose MDV has evolved to increase specific gene transcription and genome copies per cell to compensate for ICS. We speculate that the bi-directional shared pp38/origin of replication promoter, is central to this mechanism.
authors.affiliation: Hebrew University of Jerusalem, Faculty of Agriculture, Department of Animal Science, Rehovot, Israel.
authors.forename: Alon M
authors.initials: AM
authors.lastname: Levy
authors.forename: Shane C
authors.initials: SC
authors.lastname: Burgess
authors.forename: Irit
authors.initials: I
authors.lastname: Davidson
authors.forename: Greg
authors.initials: G
authors.lastname: Underwood
authors.forename: Gabriel
authors.initials: G
authors.lastname: Leitner
authors.forename: E Dan
authors.initials: ED
authors.lastname: Heller
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D002642:Chick Embryo; D017077:Culture Media, Conditioned; D016679:Genome, Viral; D008381:Herpesvirus 2, Gallid / Q000235:genetics / Q000254:growth & development / Q000378:metabolism / Q000472:pathogenicity*; D007372:Interferons / Q000494:pharmacology*; D008380:Marek Disease / Q000821:virology; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014771:Virion / Q000378:metabolism*; D014774:Virulence; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000956:Antigens, Viral; D017077:Culture Media, Conditioned; D010750:Phosphoproteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C072811:glycoprotein B, Marek's disease virus; C071698:phosphoprotein pp38, Marek's disease virus; D007372:Interferons