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Cytoskeletal changes during poliovirus infection in an intestinal cell line
Gagandeep Kang1, Prabha Desikan, Minnie Mathan
1Wellcome Trust Research Laboratory, Department of Gastrointestinal Sciences, Christian Medical College & Hospital, Vellore, India.
Background & Objectives:
Although polioviral replication has been extensively studied, cytoskeletal changes in the host cell during poliovirus replication have not been extensively investigated. We studied the ultrastructural and cytoskeletal changes in host cells during poliovirus infection.
Methods:
Fluorescence staining of filamentous actin with a fluorescein-isothiocynate labelled mycotoxin, in the absence and presence of microfilament inhibitors cytochalasins B and D, and electron microscopy were used to investigate the role and fate of actin microfilaments during poliovirus infection, morphogenesis and release in an intestinal cell line, HRT-18.
Results:
At 10 h post-infection, fluorescence staining of actin showed focal areas of fluorescence in the cytoplasm. By 16 h, these became more prominent and increased in number, and by 18-22 h they coalesced to enclose areas of the cytoplasm. These changes in the actin profile were confirmed by electron microscopy, where small actin bundles appeared in association with vesicles, increased in size, number and thickness, enclosed areas of cytoplasm with numerous vesicles and were finally seen in association with crystalline arrays of virus near the periphery of the cells. The addition of microfilament inhibitors cytochalasins B and D, after the initial period of adsorption resulted in complete inhibition of changes in the actin profile and of viral release, indicating that microfilament inhibitors prevented both polymerization of actin and movement of the virus within the cell.
Interpretation & Conclusion:
In poliovirus infection, both intracellular movement and release of virus appear to be related to cytoskeletal changes, particularly involving actin microfilaments.
Insights
Poliovirus infection alters host cell actin, impacting viral movement and release. Inhibiting actin polymerization with cytochalasins blocks these processes, revealing the role of the cytoskeleton in viral replication.
Area of Science:
- Cell Biology
- Virology
Background:
- Poliovirus replication is well-studied, but host cell cytoskeletal changes remain less understood.
- Investigating host cell ultrastructure during poliovirus infection is crucial for understanding viral mechanisms.
Purpose of the Study:
- To investigate ultrastructural and cytoskeletal alterations in host cells during poliovirus infection.
- To elucidate the role of actin microfilaments in poliovirus morphogenesis and release.
Main Methods:
- Utilized fluorescence staining of filamentous actin with a fluorescein-isothiocyanate labelled mycotoxin.
- Employed electron microscopy to examine cytoskeletal changes and viral structures.
- Assessed the impact of microfilament inhibitors (cytochalasins B and D) on actin dynamics and viral release.
Main Results:
- Observed dynamic changes in actin organization within infected cells, from focal areas to cytoplasmic enclosures.
- Electron microscopy confirmed the formation and growth of actin bundles associated with viral vesicles.
- Microfilament inhibitors completely prevented actin profile changes and viral release, indicating inhibition of actin polymerization and viral movement.
Conclusions:
- Poliovirus infection is associated with significant cytoskeletal rearrangements involving actin microfilaments.
- Intracellular movement and release of poliovirus are dependent on these actin-based cytoskeletal changes.
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