Cytoskeletal changes during poliovirus infection in an intestinal cell line

Abstract

Background and 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 and Conclusion: In poliovirus infection, both intracellular movement and release of virus appear to be related to cytoskeletal changes, particularly involving actin microfilaments.