Analysis of Lujo virus cell entry using pseudotype vesicular stomatitis virus

Area of Science:

• Virology
• Cell Biology
• Infectious Diseases

Background:

• Arenaviruses cause viral hemorrhagic fever (VHF), a significant public health concern.
• High biosafety level requirements for arenaviruses impede research and therapeutic development.
• Pseudotype viruses serve as safer surrogates for studying arenavirus biology.

Purpose of the Study:

• To develop and characterize pseudotype vesicular stomatitis virus (VSV) bearing arenavirus envelope proteins (AREpv).
• To investigate the cell entry mechanisms of Lujo virus (LUJV) pseudoparticles (LUJpv).
• To identify potential receptors and host factors involved in LUJV infection.

Main Methods:

• Generation of pseudotype VSV expressing envelope proteins from various arenaviruses, including LUJV.
• Infection assays in mammalian cell lines to assess infectivity and receptor usage.
• Treatment with lysosomotropic agents and cholesterol transport inhibitors (U18666A).
• Infection studies in Niemann-Pick C1 (NPC1)-deficient cells.

Main Results:

• Pseudotype arenaviruses (AREpv) showed high infectivity in mammalian cells.
• New World and Old World AREpv infections depended on hTfR1 and αDG, respectively.
• LUJpv infection was independent of hTfR1 and αDG, indicating a novel receptor.
• LUJpv entry is pH-dependent and requires cholesterol transport, specifically NPC1 activity.

Conclusions:

• LUJV utilizes a previously unidentified cellular receptor for entry.
• Cholesterol metabolism and NPC1 are essential for efficient LUJV pseudoparticle infection.
• These findings provide insights into LUJV pathogenesis and potential therapeutic targets.