Adult K18-hACE2 mice are suitable for studying intranasal SARS-CoV-2 infection but not direct-contact transmission

The rapid global spread of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) since 2019 emphasizes the need to understand its transmission routes, which mainly comprise airborne and contact transmission. Contact transmission, where the virus spreads through direct or indirect contact, is key to the disease epidemiology. Therefore, investigating contact transmission in animal models is crucial for understanding SARS-CoV-2 behavior and developing effective preventive measures. Although ferrets, cats, and hamsters have been established as models for studying contact transmission, the susceptibility of mice (the most commonly used experimental animal model) to SARS-CoV-2 contact infection remains uncertain. In this study, we investigated whether SARS-CoV-2 can spread via contact transmission in adult K18-hACE2 mice with different genetic backgrounds, including those with mitomycin C-induced immunodeficiency. We conducted contact-transmission experiments by co-housing K18-hACE2 mice intranasally infected with SARS-CoV-2 S type (isolated in Korea) alongside uninfected adult K18-hACE2 mice. Mice with genetically different backgrounds subjected to contact infection exhibited no changes in clinical signs or histopathological changes in the respiratory tract and extrapulmonary organs. Additionally, neither SARS-CoV-2 nor neutralizing antibodies were detected in any of the tested samples. Their immune responses remained unchanged, and contact transmission was not observed, even in immunodeficient mice. Collectively, these findings suggest that adult K18-hACE2 mice are not susceptible to contact infection with SARS-CoV-2, highlighting the role of immune mechanisms in viral spread and the limitations of this model for studying human transmission pathways. Our results underscore the importance of utilizing appropriate animal models to accurately elucidate transmission dynamics.IMPORTANCEUnderstanding the mechanisms of severe acute respiratory syndrome coronavirus-2 infection and transmission is essential for preventing and treating coronavirus disease 2019. Varying opinions exist regarding the occurrence of contact infection in mice. Here, we aimed to induce contact infection under various conditions in K18-hACE2 mice. By measuring clinical symptoms, viral loads, and neutralizing-antibody titers and conducting pathological analyses, we demonstrated that contact infection did not occur in K18-hACE2 mice. These findings underscore the importance of selecting appropriate experimental animal models to guide future studies on viral infections.