ArticleInternational journal of molecular sciences2026
Development of a Conditional Replication System Using a Lassa Virus Glycoprotein Complex-Encoding Retroviral Vector for Isolating Resistant Variants to Inhibitors in BSL-2.
Article in International journal of molecular sciences, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
A high-risk infectious disease or a Category A pathogen, Lassa virus (LASV), requires strict containment, classified as biosafety level 4 (BSL-4) conditions, which restricts research on the virus due to the scarcity of BSL-4 facilities. Thus, replication-defective pseudotyped retroviral vectors have been widely used as safe materials for neutralizing activity assays of drugs and antibodies in BSL-2. Here, we established a novel retroviral vector system encoding LASV glycoprotein complex (GPC) that can exclusively replicate in cells expressing the Gag-Pol protein of murine leukemia virus (MLV) under BSL-2 conditions. Using this conditional replication system, we successfully isolated LASV GPC variants resistant to either an anti-LASV compound, lamellarin α 20-sulfate, or a neutralizing antibody derived from a Lassa fever survivor. In the lamellarin α 20-sulfate-resistant variants, K125E and H13R amino acid substitutions cooperatively conferred resistance. The K125E enhanced infectivity and simultaneously conferred a lethal effect on cells in the conditional replication system, while the H13R mitigated the latter effect, thereby enabling stable expression of LASV GPC in cells. In the neutralizing antibody-resistant variants, I403T substitution was responsible for the resistance by impairing antibody binding. This study provides a valuable BSL-2-based platform for isolating LASV GPC variants resistant to inhibitors and characterizing their mutations.
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