Lassa virus (LASV) is recognized by the World Health Organization as one of the top five pathogens likely to cause a severe outbreak. A recent unprecedented resurgence of LASV in Nigeria caused by genetically diverse strains underscores the need for licensed medical countermeasures. Single-injection vaccines that can rapidly control outbreaks and confer long-term immunity are needed. Vaccination of cynomolgus monkeys with a recombinant vesicular stomatitis virus vector expressing the glycoprotein precursor of LASV lineage IV strain Josiah (rVSVΔG-LASV-GPC) induces fast-acting protection in monkeys challenged 3 or 7?days later with a genetically heterologous lineage II isolate of LASV from Nigeria, while nonspecifically vaccinated control animals succumb to challenge. The rVSVΔG-LASV-GPC vaccine induces rapid activation of adaptive immunity and the transcription of natural killer (NK) cell-affiliated mRNAs. This study demonstrates that rVSVΔG-LASV-GPC may provide rapid protection in humans against LASV infections in cases where immediate public-health intervention is required.