De novo recovery of Ghana virus, an African bat Henipavirus, reveals differential tropism and attenuated pathogenicity compared to Nipah virus

Henipaviruses (HNVs) like Nipah (NiV) and Hendra (HeV) viruses represent potent zoonotic threats. Ghana virus (GhV), identified through metagenomics in 2012, is the only African bat-borne henipavirus with a near full-length sequenced genome. However, because GhV has not been isolated in culture, its biology, pathogenicity, and zoonotic potential remain poorly understood. Using reverse genetics at BSL-4, we recovered a full-length infectious clone of recombinant GhV, requiring rational reconstruction of the incomplete 3' leader and correction of a non-canonical transcriptional initiation site. GhV showed restricted receptor tropism (ephrin-B2 but not ephrin-B3) and distinct innate immune antagonism mechanisms compared to NiV. Relative to NiV, GhV replication was attenuated in primary human and porcine cells but enhanced in specific bat cell lines. Experimental infection of Syrian golden hamsters with GhV resulted in no observable disease or mortality. Furthermore, a chimeric NiV encoding the GhV receptor binding protein was completely attenuated in vivo, strongly implicating ephrin-B3 receptor usage as a key determinant of henipavirus pathogenesis. We further identified several broad-spectrum antiviral compounds that inhibit Ghana virus replication. Our studies illuminate critical determinants of HNV pathogenicity and replication, enhance our understanding of GhV zoonotic potential, and inform future strategies for virus surveillance and control.