ABSTRACT
Lujo virus (LUJV), a highly pathogenic arenavirus, was first identified in 2008 in Zambia. To aid the identification of effective therapeutics for LUJV, we developed a recombinant reporter virus system, confirming reporter LUJV comparability with wild-type virus and its utility in high-throughput antiviral screening assays. Using this system, we evaluated compounds with known and unknown efficacy against related arenaviruses, with the aim of identifying LUJV-specific and potential new pan-arenavirus antivirals. We identified six compounds demonstrating robust anti-LUJV activity, including several compounds with previously reported activity against other arenaviruses. These data provide critical evidence for developing broad-spectrum antivirals against high-consequence arenaviruses.
Subject(s)
Antiviral Agents/pharmacology , Arenavirus/drug effects , Lujo virus/drug effects , Animals , Arenaviridae Infections/drug therapy , Arenaviridae Infections/virology , Arenavirus/physiology , Cell Line, Tumor , Chlorocebus aethiops , Genome, Viral , Green Fluorescent Proteins/genetics , Humans , Lujo virus/genetics , Lujo virus/physiology , Microbial Sensitivity Tests , Recombinant Proteins , Vero Cells , Virus Internalization/drug effectsABSTRACT
Arenaviruses cause fatal hemorrhagic disease in humans. Old World arenavirus glycoproteins (GPs) mainly engage α-dystroglycan as a cell-surface receptor, while New World arenaviruses hijack transferrin receptor. However, the Lujo virus (LUJV) GP does not cluster with New or Old World arenaviruses. Using a recombinant vesicular stomatitis virus containing LUJV GP as its sole attachment and fusion protein (VSV-LUJV), we demonstrate that infection is independent of known arenavirus receptor genes. A genome-wide haploid genetic screen identified the transmembrane protein neuropilin 2 (NRP2) and tetraspanin CD63 as factors for LUJV GP-mediated infection. LUJV GP binds the N-terminal domain of NRP2, while CD63 stimulates pH-activated LUJV GP-mediated membrane fusion. Overexpression of NRP2 or its N-terminal domain enhances VSV-LUJV infection, and cells lacking NRP2 are deficient in wild-type LUJV infection. These findings uncover this distinct set of host cell entry factors in LUJV infection and are attractive focus points for therapeutic intervention.
Subject(s)
Lujo virus/physiology , Neuropilin-2/metabolism , Tetraspanin 30/metabolism , Viral Fusion Proteins/metabolism , Viral Proteins/metabolism , Virus Internalization , Carrier Proteins , Cell Line , Host-Pathogen Interactions/physiology , Human Umbilical Vein Endothelial Cells , Humans , Lujo virus/genetics , Lujo virus/pathogenicity , Protein Interaction Domains and Motifs , Receptors, Cell Surface/metabolism , Receptors, Transferrin , Viral Fusion Proteins/genetics , Viral Proteins/geneticsABSTRACT
Lujo virus is an emerging arenavirus circulating in Southern Africa. Although to date there has only been a single outbreak of the novel haemorrhagic disease resulting from human infection with this virus, the case-fatality rate of exposed individuals, including nosocomial transmission, was 80%. The ability to identify viral haemorrhagic fevers accurately, especially those capable of nosocomial transmission, is of critical importance. Timely identification of these diseases allow medical professionals to isolate patients and implement barrier nursing techniques in order to prevent onward transmission of the virus. While rapid diagnostic methods are published for most viral haemorrhagic fevers, at present there are no such virus specific protocols for Lujo haemorrhagic fever. This report details the first set of diagnostic molecular assays designed to identify Lujo viral RNA rapidly, and demonstrates the potential functionality of these assays for use in the clinical setting. Although these assays have been designed and validated against a solitary isolate of Lujo virus, this represents the entirety of strains detected to date, and offer quick, cheap and easy methods for use in diagnostic laboratories.