Recombinant Lloviu virus as a tool to study viral replication and host responses

PLoS Pathog. 2022 Feb 4;18(2):e1010268. doi: 10.1371/journal.ppat.1010268. eCollection 2022 Feb.

Abstract

Next generation sequencing has revealed the presence of numerous RNA viruses in animal reservoir hosts, including many closely related to known human pathogens. Despite their zoonotic potential, most of these viruses remain understudied due to not yet being cultured. While reverse genetic systems can facilitate virus rescue, this is often hindered by missing viral genome ends. A prime example is Lloviu virus (LLOV), an uncultured filovirus that is closely related to the highly pathogenic Ebola virus. Using minigenome systems, we complemented the missing LLOV genomic ends and identified cis-acting elements required for LLOV replication that were lacking in the published sequence. We leveraged these data to generate recombinant full-length LLOV clones and rescue infectious virus. Similar to other filoviruses, recombinant LLOV (rLLOV) forms filamentous virions and induces the formation of characteristic inclusions in the cytoplasm of the infected cells, as shown by electron microscopy. Known target cells of Ebola virus, including macrophages and hepatocytes, are permissive to rLLOV infection, suggesting that humans could be potential hosts. However, inflammatory responses in human macrophages, a hallmark of Ebola virus disease, are not induced by rLLOV. Additional tropism testing identified pneumocytes as capable of robust rLLOV and Ebola virus infection. We also used rLLOV to test antivirals targeting multiple facets of the replication cycle. Rescue of uncultured viruses of pathogenic concern represents a valuable tool in our arsenal for pandemic preparedness.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cell Line
  • Chlorocebus aethiops
  • Ebolavirus / genetics*
  • Filoviridae / genetics*
  • Filoviridae Infections / virology*
  • Genetic Complementation Test
  • Genome, Viral
  • Hemorrhagic Fever, Ebola / virology
  • Host Microbial Interactions
  • Humans
  • Inclusion Bodies / virology
  • Induced Pluripotent Stem Cells / virology
  • Macrophages / virology
  • RNA, Viral
  • Reverse Genetics
  • Vero Cells
  • Virion / genetics
  • Virus Replication*

Substances

  • RNA, Viral