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Early Entry Events in Echovirus 30 Infection.

Helena Vandesande1, Mira Laajala2, Tino Kantoluoto2

  • 1Department of Chemistry and Biomedical Sciences, Linnaeus University, Kalmar, Sweden.

Journal of Virology
|April 17, 2020
PubMed
Summary

Echovirus 30 (E30) uses the Fc echovirus receptor (FcRn) for cell entry and follows a macropinocytic pathway, not clathrin-mediated endocytosis, to infect human cells. This study clarifies the early entry and replication of E30, a common cause of viral meningitis.

Keywords:
aseptic meningitisearly entryechovirus 30enterovirus

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Area of Science:

  • Virology
  • Cell Biology
  • Molecular Biology

Background:

  • Echovirus 30 (E30) is a significant cause of viral meningitis in children and adults, responsible for global outbreaks.
  • Despite its prevalence, the precise mechanisms of E30 cell entry and replication remain largely uncharacterized.
  • Understanding E30's infectious pathway is crucial for developing targeted therapeutic strategies.

Purpose of the Study:

  • To elucidate the early entry and replication events of Echovirus 30 (E30) in human RD cells.
  • To identify the specific cellular receptors and pathways utilized by E30 during infection.
  • To compare E30's entry mechanism with other known enteroviruses.

Main Methods:

  • Generation of Echovirus 30 strain Bastianni (E30B) from an infectious cDNA clone.
  • Infection assays in human RD cells using E30B, assessing the roles of Fc echovirus receptor (FcRn), CAR, and DAF.
  • Pharmacological inhibition of cellular pathways (Rac1, cholesterol, macropinocytosis, clathrin-mediated endocytosis) and analysis of viral replication intermediates and capsid production.
  • Colocalization studies with early endosomes and assessment of Rab5 expression effects on infection.

Main Results:

  • E30B infection requires FcRn but not CAR; interaction with DAF was observed but not essential for entry.
  • Viral RNA replication initiated between 2-3 hours post-infection, and capsid production began 4-5 hours post-infection.
  • Infection was inhibited by drugs targeting Rac1 and cholesterol, suggesting a macropinocytic entry pathway, distinct from clathrin-mediated endocytosis.
  • E30B colocalized with early endosomes, and Rab5 expression enhanced infection, indicating partial overlap with classical early endosomes.

Conclusions:

  • Echovirus 30 utilizes a unique entry pathway involving FcRn and macropinocytosis, leading to uncoating within early endosomes.
  • E30 does not rely on acidification for entry and bypasses the clathrin-mediated endocytosis pathway.
  • The findings contribute to defining a conserved 'enterovirus entry pathway' within the Enterovirus B species.