Comparison of three pangolin coronaviruses showing distinct pathogenicity, transmissibility and potential for cross-species infection

Pangolins harbor SARS-CoV-2–related coronaviruses (PCoVs). We isolate three PCoVs (P8V, P11V, P5V) from confiscated Malayan pangolins and analyzed their genomes, evolution and functional properties. Phylogenetic analyses place these viruses in two clusters within the PCoV-GX lineage, revealing spike mutations that later convergently evolve in SARS-CoV-2. All viruses replicate efficiently in Vero E6 cells, with variable replication in human cell lines and organoids. In hACE2-transgenic mice, P11V and previously isolated P2V cause rapid, severe disease, whereas P8V and P5V exhibit delayed pathogenicity. Direct-contact transmission in hamsters is efficient for all strains, with lung pathology reflecting strain-specific virulence. These results demonstrate that PCoVs possess substantial infectivity, pathogenicity, and transmissibility, illustrating their adaptability, and highlight the role of pangolins as reservoirs of coronaviruses with zoonotic potential. Comparative analysis of three pangolin coronaviruses reveals distinct pathogenicity and transmissibility profiles, demonstrates their potential for cross species infection, and highlights pangolins as potential reservoirs of zoonotic coronavirus threats.