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Assays for the Specific Growth Rate and Cell-binding Ability of Rotavirus
Published on: January 28, 2019
Rotavirus-Mediated Prostaglandin E2 Production in MA104 Cells Promotes Virus Attachment and Internalisation,
Willem J Sander1, Gabré Kemp1, Arnold Hugo2
1Department of Microbiology and Biochemistry, University of the Free State, Bloemfontein, South Africa.
Insights
Rotavirus infection increases prostaglandin E2 production, which aids viral entry into cells. Inhibiting this molecule reduces rotavirus yield, suggesting it as a potential antiviral target.
Area of Science:
- Virology
- Cell Biology
- Immunology
Background:
- Rotaviruses cause severe diarrhea in young children globally.
- Existing vaccines have limited impact in sub-Saharan Africa, and no antivirals are available.
- Prostaglandin E2 (PGE2) is produced during rotavirus infection and influences disease progression.
Purpose of the Study:
- To investigate the role of prostaglandin E2 (PGE2) in rotavirus infection.
- To explore the potential of targeting PGE2 biosynthesis as an antiviral strategy.
Main Methods:
- MA104 cells were supplemented with gamma-linolenic acid (GLA) and infected with rotavirus SA11.
- Prostaglandin E2 levels were measured using ELISA.
- Confocal microscopy was used to visualize PGE2 co-localization with viral proteins.
- Replication kinetics and flow cytometry were employed to assess viral yield and entry.
Main Results:
- Rotavirus infection depleted GLA and arachidonic acid, increasing PGE2 production.
- PGE2 co-localized with viroplasm proteins (NSP5, NSP2), suggesting a role for viroplasms in PGE2 production.
- Inhibitors of PGE2 synthesis reduced rotavirus yield, particularly in early infection stages.
- PGE2 enhanced rotavirus attachment and internalization, facilitating clathrin-mediated entry.
Conclusions:
- Prostaglandin E2 plays a crucial role in enhancing rotavirus attachment and entry into host cells.
- Viroplasms may be involved in the production of rotavirus-induced PGE2.
- Targeting prostaglandin E2 biosynthesis presents a promising avenue for developing novel antiviral therapies against rotavirus.
Abstract:
Rotaviruses are one of the leading causes of severe dehydrating diarrhoea in infants and children under the age of five. Despite the introduction of vaccines, disease burden remains high in sub-Saharan Africa, with no known anti-viral treatments available. During early infection rotavirus attaches to several cellular receptors and enters the cells by either clathrin-dependent or -independent endocytosis. Prostaglandin E2, an abundant eicosanoid, is produced from arachidonic acid during rotavirus infection and inhibition of prostaglandin E2 formation have a deleterious effect on rotavirus infection. In this study, MA104 cells were supplemented with γ-linolenic acid (GLA), a precursor of arachidonic acid. Infection of supplemented cells with rotavirus SA11 led to a depletion in the relative percentages of GLA and arachidonic acid which coincided with an increased production of prostaglandin E2 as monitored by ELISA. Confocal microscopy demonstrated that prostaglandin E2 co-localises with the viroplasm-forming proteins, NSP5 and NSP2. Due to the known association of viroplasms with lipid droplets and the fact that lipid droplets are sites for prostaglandin E2 production, our results indicate a possible role for viroplasms in the production of rotavirus-induced prostaglandin E2. Replication kinetics showed that inhibitors, targeting the biosynthesis of prostaglandin E2, had negative effects on rotavirus yield, especially during the early stages of infection. Using flow cytometry and prostaglandin E2 addback experiments, we show that prostaglandin E2 enhances the attachment and internalisation of rotavirus in MA104 cells indicating a possible role for prostaglandin E2 during clathrin-mediated rotavirus entry. The production of prostaglandin E2 during rotavirus infection could serve as a possible target for anti-viral treatment.
