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Infection of liver sinusoidal endothelial cells with Muromegalovirus muridbeta1 involves binding to neuropilin-1 and

Ingelin Kyrrestad1, Anett Kristin Larsen1, Javier Sánchez Romano1

  • 1Department of Medical Biology, Faculty of Health Sciences, UiT - The Arctic University of Norway, Tromsø, Norway.

Frontiers in Cellular and Infection Microbiology
|November 29, 2023
PubMed

Insights

Murine cytomegalovirus (MuHV-1) infection of liver sinusoidal endothelial cells (LSECs) depends on dynamin, a protein crucial for endocytosis. This suggests MuHV-1 enters LSECs via dynamin-mediated endocytosis, potentially involving neuropilin-1 binding.

Area of Science:

  • Virology
  • Cell Biology
  • Immunology

Background:

  • Liver sinusoidal endothelial cells (LSECs) are critical scavenger cells involved in macromolecule and nanoparticle clearance.
  • LSECs utilize clathrin-coated pit-mediated endocytosis, a dynamin-dependent process, for cellular uptake.
  • The entry mechanism of betaherpesviruses, including MuHV-1, into LSECs remains largely undescribed.

Purpose of the Study:

  • To investigate the role of dynamin in the early stages of MuHV-1 infection in primary mouse LSECs.
  • To establish a reliable protocol for dynamin inhibition studies in LSEC cultures.
  • To elucidate the specific endocytic pathway utilized by MuHV-1 for LSEC entry.

Main Methods:

  • Primary mouse LSEC cultures were established and treated with dynamin inhibitors (dynasore, MitMAB) to assess effects on cell viability and endocytosis.
  • LSECs were infected with MuHV-1 in the presence or absence of dynamin inhibitors, monensin, or a neuropilin-1 antibody.
  • Viral infection was quantified by measuring nuclear immediate early antigen (IE1) expression via immunofluorescence.

Main Results:

  • MuHV-1 efficiently infected primary mouse LSECs in vitro.
  • Dynasore and MitMAB significantly and independently inhibited MuHV-1 infection, indicating a dynamin-dependent entry process.
  • Monensin treatment reduced infection, suggesting endosomal acidification is involved, while neuropilin-1 antibody binding blocked LSEC infection, pointing to its role in viral attachment.

Conclusions:

  • The initial steps of MuHV-1 infection in mouse LSECs are dynamin-dependent.
  • MuHV-1 likely enters LSECs via endocytosis, involving binding to neuropilin-1.
  • These findings provide crucial insights into betaherpesvirus tropism and entry mechanisms into liver sinusoidal endothelial cells.
Keywords:
CMVLSECMitMABMuHV-1cytomegalovirusdynasoreendocytosisneuropilin-1

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