Significance Middle East respiratory syndrome, caused by a zoonotically transmitted coronavirus (MERS-CoV), has a high mortality (∼36%). Because of limited autopsy data on tissues from MERS fatalities, a small animal model can provide an important tool to better understand the disease. We humanized the mouse locus of the virus receptor DPP4, preserving native DPP4 expression. After inoculating hDPP4 knockin mice with MERS-CoV, there was virus replication without disease. We then generated a mouse-adapted MERS-CoV by serial passage in hDPP4 knockin mice. The resultant virus causes fatal lung disease that includes diffuse alveolar damage and immune dysregulation. Here, we characterize the pathologic features of the model and elucidate key aspects of the immunopathology and factors contributing to virulence. The Middle East respiratory syndrome (MERS) emerged in Saudi Arabia in 2012, caused by a zoonotically transmitted coronavirus (CoV). Over 1,900 cases have been reported to date, with ∼36% fatality rate. Lack of autopsies from MERS cases has hindered understanding of MERS-CoV pathogenesis. A small animal model that develops progressive pulmonary manifestations when infected with MERS-CoV would advance the field. As mice are restricted to infection at the level of DPP4, the MERS-CoV receptor, we generated mice with humanized exons 10–12 of the mouse Dpp4 locus. Upon inoculation with MERS-CoV, human DPP4 knockin (KI) mice supported virus replication in the lungs, but developed no illness. After 30 serial passages through the lungs of KI mice, a mouse-adapted virus emerged (MERS MA ) that grew in lungs to over 100 times higher titers than the starting virus. A plaque-purified MERS MA clone caused weight loss and fatal infection. Virus antigen was observed in airway epithelia, pneumocytes, and macrophages. Pathologic findings included diffuse alveolar damage with pulmonary edema and hyaline membrane formation associated with accumulation of activated inflammatory monocyte–macrophages and neutrophils in the lungs. Relative to the parental MERS-CoV, MERS MA viruses contained 13–22 mutations, including several within the spike (S) glycoprotein gene. S-protein mutations sensitized viruses to entry-activating serine proteases and conferred more rapid entry kinetics. Recombinant MERS MA bearing mutant S proteins were more virulent than the parental virus in hDPP4 KI mice. The hDPP4 KI mouse and the MERS MA provide tools to investigate disease causes and develop new therapies.
【저자키워드】 interferon, Spike protein, Emerging pathogen, CD26, virus pathogenesis,