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[Accumulation of certain arenaviruses in transplantable VERO and BHK-21 cells]
Abstract:
Production of Lassa and Machupo viruses in Vero and BHK-21 cells was studied in relation to various conditions of the infected cell cultivation and as a function of different multiplicities of infection. The highest titers (expressed in PFU/ml) were obtained when the cells were grown in roller bottles with daily changes of the medium. The maximum titer in Lassa virus-infected cells was over 10(6), in Machupo virus-infected cells over 10(7). The effect of the autointerfering factor on the growth of Machupo virus was demonstrated. An increase in the multiplicity of infection led to a decrease in the yield of Machupo virus. Production of Pichinde and Machupo viruses in a monocyclic growth experiment was studied. The maximum yield of cell-associated and extracellular Pichinde virus was obtained at 24-32 hours postinfection, and that of extracellular Machupo virus at 32-40 hours postinfection.
Insights
Optimizing cell culture conditions, such as daily medium changes in roller bottles, significantly enhances Lassa and Machupo virus production. Higher multiplicities of infection decreased Machupo virus yield due to autointerference.
Area of Science:
- Virology
- Cell Biology
Context:
- Lassa and Machupo viruses are significant human pathogens requiring efficient laboratory production for research.
- Optimizing viral production in cell culture is crucial for diagnostic and therapeutic development.
Purpose:
- To investigate the optimal conditions for producing Lassa, Machupo, and Pichinde viruses in cell cultures.
- To evaluate the impact of cultivation methods and multiplicity of infection on viral yield.
Summary:
- Highest titers of Lassa and Machupo viruses were achieved in Vero and BHK-21 cells using roller bottles with daily medium changes, reaching over 10^6 PFU/ml for Lassa and 10^7 PFU/ml for Machupo.
- Increased multiplicity of infection negatively impacted Machupo virus yield, indicating an autointerfering factor.
- Pichinde virus production peaked at 24-32 hours post-infection, while extracellular Machupo virus yield was maximal at 32-40 hours.
Impact:
- Provides critical insights into optimizing viral propagation for Lassa, Machupo, and Pichinde viruses.
- Informs the development of improved cell culture protocols for arenavirus research and diagnostics.