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Dissecting Host-virus Interaction in Lytic Replication of a Model Herpesvirus
Published on: October 7, 2011
Biological and macromolecular properties of murine cells persistently infected with MHV-JHM
Abstract:
A persistently-infected neuroblastoma culture [Neuro-2A( JHMV )] was established with the murine hepatitis virus JHM [MHV-JHM]. After 100 days of passage, the endogenous virus [Neuro-2A( JHMV ) end] released by this culture was unable to induce the syncytia typical of MHV-JHM and the endogenous virus was not temperature-sensitive. The Neuro-2A( JHMV ) culture was cured of virus production by passage under neutralizing antibody [Neuro-2A( JHMV )Ab]. The Neuro-2A( JHMV ) and the Neuro-2A ( JHMV ) Ab cultures were as susceptible to heterologous infection with mengovirus and vesicular stomatitis virus as the uninfected Neuro-2A culture. However, the Neuro-2A ( JHMV ) and Neuro-2A( JHMV ) Ab cultures were partially resistant to homologous superinfection by MHV-JHM and the closely related MHV-A59. Virus related to MHV-JHM was rescued from the antibody-cured cells by cell fusion. The synthesis of MHV-JHM specific antigens by Neuro-2A( JHMV ) cells, Neuro-2A( JHMV ) Ab cells and 17 Cl-1 cells infected by Neuro-2A( JHMV ) end was studied by SDS-PAGE. The genomic RNAs of MHV-JHM and Neuro-2A( JHMV ) end were compared by oligonucleotide mapping. The results of the protein and RNA studies indicated that the genome of Neuro-2A( JHMV ) end was substantially modified from the genome of MHV-JHM, but the modifications did not significantly alter the molecular size of the viral-specific proteins.
Insights
Persistent murine hepatitis virus JHM (MHV-JHM) infection in neuroblastoma cells resulted in an endogenous virus with a modified genome. This altered virus exhibited resistance to superinfection but retained susceptibility to other viruses.
Area of Science:
- Virology
- Cell Biology
- Neuroscience
Background:
- Persistent viral infections can lead to viral evolution and altered pathogenesis.
- Murine hepatitis virus JHM (MHV-JHM) is a coronavirus known to cause persistent infections in the central nervous system.
Purpose of the Study:
- To characterize an endogenous virus derived from a persistently infected neuroblastoma cell line (Neuro-2A).
- To investigate the genetic and phenotypic changes in MHV-JHM following prolonged in vitro culture.
Main Methods:
- Establishment of a persistently infected neuroblastoma cell culture (Neuro-2A(JHMV)).
- Viral characterization through syncytia induction, temperature sensitivity assays, and heterologous/homologous superinfection studies.
- Analysis of viral antigens using SDS-PAGE and genomic RNA comparison via oligonucleotide mapping.
Main Results:
- An endogenous virus (Neuro-2A(JHMV)end) was released, lacking syncytia-inducing and temperature-sensitive properties of the parent MHV-JHM.
- Infected and cured cell lines showed partial resistance to homologous MHV-JHM and MHV-A59 superinfection.
- Oligonucleotide mapping and protein analysis revealed substantial genomic modification in the endogenous virus, without significant alteration in viral protein size.
Conclusions:
- Prolonged MHV-JHM infection in Neuro-2A cells leads to viral evolution with significant genomic alterations.
- The modified endogenous virus exhibits altered biological properties, including resistance to superinfection, suggesting viral adaptation during persistent infection.
