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Arenavirus defective interfering particles mask the cell-killing potential of standard virus
Abstract:
Lymphocytic choriomeningitis virus (LCM) and Pichinde virus grew readily and produced cytopathology in MDCK and PK-15 cells. It is known that in these cell lines, the synthesis or function of defective interfering (DI) virus particles is restricted. Survival curves of single MDCK cells infected with low multiplicities of LCM showed one-particle-to-kill kinetics. At high multiplicities of infection, there was a maximum degree of cell-killing, or even a reduction in the amount of cell-killing, depending on how much DI virus was present in a particular standard virus stock. DI LCM virus could completely prevent standard virus from producing c.p.e. in MDCK monolayers with one-particle-to-protect kinetics. It could still prevent killing of the cells when added within a short time after infection with standard virus, but was able to interfere with synthesis of standard virus when added even later, On passage of LCM or Pichinde virus without dilution in MDCK cells, there was no homologous auto-interference. Furthermore, there was only slight interference with the synthesis of standard virus when these cells were pre-treated with DI virus.
Insights
Defective interfering (DI) Lymphocytic choriomeningitis virus (LCM) particles restrict standard LCM virus from causing cell damage in cell cultures. DI virus can prevent cell killing and interfere with standard virus replication.
Area of Science:
- Virology
- Cell Biology
Background:
- Lymphocytic choriomeningitis virus (LCM) and Pichinde virus replicate and cause cytopathology in MDCK and PK-15 cells.
- Defective interfering (DI) virus particle synthesis or function is restricted in these cell lines.
Purpose of the Study:
- To investigate the role of DI virus particles in LCM virus infection dynamics.
- To determine the kinetics of cell killing and protection by standard and DI LCM virus.
Main Methods:
- Infection of MDCK cells with varying multiplicities of standard and DI LCM virus.
- Analysis of cell survival curves and cytopathic effect (c.p.e.).
- Assessment of interference with standard virus synthesis.
Main Results:
- Single-cell infections with low multiplicities of LCM showed one-particle-to-kill kinetics.
- High multiplicities of LCM resulted in maximum cell killing, or reduced killing depending on DI virus presence.
- DI LCM virus demonstrated one-particle-to-protect kinetics, preventing c.p.e. and interfering with standard virus synthesis.
- No homologous auto-interference was observed upon undiluted passage of LCM or Pichinde virus.
Conclusions:
- DI LCM virus particles play a significant role in modulating LCM virus pathogenesis in cell culture.
- DI virus can effectively interfere with standard virus replication and prevent cell death, even when introduced post-infection.
- The restricted synthesis of DI particles in MDCK cells influences the outcome of LCM virus infection.