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Persistent infection of K562 cells by encephalomyocarditis virus
I U Pardoe1, K K Grewal, M P Baldeh
1Faculty of Medicine, Memorial University of Newfoundland, St. John's, Canada.
Abstract:
Infection of human erythroleukemic K562 cells by encephalomyocarditis virus readily resulted in establishment of persistently infected cultures. In contrast to the usual typical lytic infection by encephalomyocarditis virus, in which trypan blue staining of cells reaches close to 100% by about 15 h postinfection, K562 cell cultures required 3 to 4 days postinfection to reach a maximum of about 80 to 90% cell staining. The proportion of K562 cells taking up stain gradually decreased to about 10% of those present by about 13 days postinfection; during this time, virus yield per day measured by either plaque or hemagglutination titration fell about 10-fold. The decrease in percent staining was followed by waves of increased staining accompanied by increased virus production. Virus-producing cultures were maintained for over 3 months. Evolution of both virus and cells accompanied establishment of persistence in that plaque size changed from about 7 mm in diameter for the original virus to less than 1.5 mm by day 20 postinfection and most of the cells cloned from persistently infected cultures were resistant to superinfection with the original virus. Resistance was due, at least in part, to reduced virus attachment in that binding of 3H-labeled virus to cloned resistant cells was about 2% of that to uninfected cells.
Insights
Encephalomyocarditis virus establishes persistent infections in K562 cells, unlike typical lytic infections. Evolved viruses show reduced plaque size and K562 cells develop resistance to superinfection, partly due to decreased virus attachment.
Area of Science:
- Virology
- Cell Biology
- Immunology
Background:
- Encephalomyocarditis virus (EMCV) typically causes lytic infections in susceptible cells.
- Persistent viral infections can arise from complex interactions between virus and host cells.
- K562 cells are a human erythroleukemic cell line often used in virological studies.
Purpose of the Study:
- To investigate the establishment and characteristics of persistent encephalomyocarditis virus infections in K562 cells.
- To characterize the viral and cellular evolution during persistent infection.
- To determine the mechanisms underlying K562 cell resistance to superinfection.
Main Methods:
- Infection of K562 cells with EMCV to establish persistent cultures.
- Monitoring of cell viability (trypan blue staining) and virus production (plaque assay, hemagglutination titration) over time.
- Characterization of viral evolution through plaque size measurements.
- Cloning of persistently infected cells and assessment of superinfection resistance and viral attachment.
Main Results:
- EMCV established persistent infection in K562 cells, with delayed and incomplete initial cell lysis.
- Virus yield and cell staining fluctuated, with waves of increased virus production observed.
- Over 3 months, the virus evolved, showing a significant decrease in plaque size.
- Cloned K562 cells from persistent cultures exhibited resistance to superinfection, attributed partly to reduced viral attachment.
Conclusions:
- K562 cells can support persistent EMCV infection, deviating from the typical lytic pathway.
- Viral and cellular evolution, including reduced plaque size and superinfection resistance, are key features of this persistent infection model.
- Reduced virus attachment is a significant factor contributing to K562 cell resistance to superinfection in persistently infected cultures.
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