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Recovery of structurally intact and infectious poliovirus type 1 from HeLa cells during receptor-mediated endocytosis

P Willingmann1, H Barnert, H Zeichhardt

  • 1Institut für Klinische und Experimentelle Virologie, Freie Universität Berlin, Federal Republic of Germany.

Virology
|February 1, 1989
PubMed

Insights

Poliovirus type 1 enters human cells via receptor-mediated endocytosis, remaining intact and infectious for up to 30 minutes. Viral uncoating occurs later in acidic endosomes, suggesting a role for clathrin-coated vesicles in poliovirus entry.

Area of Science:

  • Virology
  • Cell Biology
  • Molecular Biology

Background:

  • Poliovirus is a significant human pathogen.
  • Understanding viral entry mechanisms is crucial for developing antiviral strategies.

Purpose of the Study:

  • To investigate the entry pathway of poliovirus type 1 into HeLa cells.
  • To determine the integrity and infectivity of poliovirus during the initial stages of cellular entry.
  • To elucidate the role of specific cellular structures in poliovirus endocytosis.

Main Methods:

  • Recovery of intact poliovirus particles from infected HeLa cells at various time points post-adsorption.
  • Characterization of recovered virus particles for integrity (sedimentation analysis, VP4 content) and infectivity (N-antigenicity).
  • Localization of virus particles within cellular compartments using microscopy.
  • Treatment of endocytic vesicles with Tris buffer to assess viral release.

Main Results:

  • Intact, infectious poliovirus type 1 (156 S particles containing VP4) was recovered from HeLa cells up to 30 minutes after adsorption.
  • A decrease in intact, infectious virus recovery was observed after 30 minutes, correlating with virus localization in acidic endosomes/lysosomes.
  • Poliovirus was found enclosed in clathrin-coated vesicles at 10 minutes post-adsorption.
  • Disruption of these coated vesicles with Tris buffer was necessary to recover intact, infectious virus.

Conclusions:

  • Poliovirus type 1 enters HeLa cells intact via receptor-mediated endocytosis, utilizing clathrin-coated structures.
  • The virus remains protected and infectious within endocytic vesicles for at least 30 minutes.
  • Viral uncoating, leading to loss of infectivity, likely occurs within acidic endosomal or lysosomal compartments.

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