Transcriptome analysis in air–liquid interface porcine respiratory epithelial cell cultures reveals that the betacoronavirus porcine encephalomyelitis hemagglutinating virus induces a robust interferon response to infection

Authors: SARLO DAVILA, KAITLYN - Oak Ridge Institute For Science And Education (ORISE); NELLI, RAHUL - Iowa State University; MORA-DIAZ, JUAN CARLOS - Iowa State University; SANG, YONGMING - Tennessee State University; Miller, Laura; GIMENEZ-LIROLA, LUIS - Iowa State University

Submitted to: Viruses
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/7/2024
Publication Date: 6/11/2024
Citation: Sarlo Davila, K.M., Nelli, R.K., Mora-Diaz, J., Sang, Y., Miller, L.C., Gimenez-Lirola, L.G. 2024. Transcriptome analysis in air–liquid interface porcine respiratory epithelial cell cultures reveals that the betacoronavirus porcine encephalomyelitis hemagglutinating virus induces a robust interferon response to infection. Viruses. https://doi.org/10.3390/v16060939.
DOI: https://doi.org/10.3390/v16060939

Interpretive Summary: This study is a continuation of our previous studies with air-liquid interface porcine respiratory epithelial cells (ALI-PRECs) inoculated with the neurotropic betacoronavirus porcine hemagglutinating encephalomyelitis virus (PHEV) which primarily infects and replicates in the upper respiratory tract of swine. The neurotropic beta coronavirus porcine hemagglutinating encephalomyelitis virus (PHEV) is an under-researched coronavirus associated with respiratory disease in pigs. This study investigated the mechanisms modulating the host response to PHEV infection over time using air-liquid interface porcine respiratory cell cultures. The results of this study shed light on the mechanisms driving the immune response to PHEV, especially the interferon and chemokine responses, involved in antiviral regulation against PHEV. The approach shown here serves as a virus-animal model to study prevalent coronaviral infections.

Technical Abstract: Primary porcine respiratory epithelial cells (PRECs) were cultured in an air-liquid interface (ALI) to differentiate into a pseudostratified columnar epithelium, proliferative basal cells, M cells, ciliated cells, and mucus-secreting goblet cells recreating a cell culture environment morphologically and functionally more representative of the epithelial lining of the swine trachea than traditional culture systems. ALI-PRECs were infected with porcine hemagglutinating encephalomyelitis virus (PHEV) which replicated actively in this environment, inducing cytopathic changes and progressive disruption of the mucociliary apparatus. The innate immunity against PHEV was evaluated in ALI-PREC cultures via transcriptomic analysis at 24, 36, and 48hpi. We showed that antiviral signaling pathways were significantly activated to mediate increased expression of antiviral IFN-stimulated genes including RSAD2, MX1, IFIT, and ISG15, as well as chemokine genes such as CCL5 and CXCL10 underlying inflammatory regulation. This study sheds light on the molecular mechanisms driving the innate immune response to PHEV at the airway epithelium, underscoring the important role of the interferon and chemokine response to PHEV.