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Our previous studies showed that bovine respiratory syncytial virus (BRSV) followed by  Histophilus somni  causes more severe bovine respiratory disease and a more permeable alveolar barrier  in vitro  than either agent alone. However, microarray analysis revealed the treatment of bovine alveolar type 2 (BAT2) epithelial cells with  H .  somni  concentrated culture supernatant (CCS) stimulated up-regulation of four antiviral protein genes as compared with BRSV infection or dual treatment. This suggested that inhibition of viral infection, rather than synergy, may occur if the bacterial infection occurred before the viral infection. Viperin (or radical S-adenosyl methionine domain containing 2—RSAD2) and ISG15 (IFN-stimulated gene 15—ubiquitin-like modifier) were most up-regulated. CCS dose and time course for up-regulation of viperin protein levels were determined in treated bovine turbinate (BT) upper respiratory cells and BAT2 lower respiratory cells by Western blotting. Treatment of BAT2 cells with  H .  somni  culture supernatant before BRSV infection dramatically reduced viral replication as determined by qRT PCR, supporting the hypothesis that the bacterial infection may inhibit viral infection. Studies of the role of the two known  H .  somni  cytotoxins showed that viperin protein expression was induced by endotoxin (lipooligosaccharide) but not by IbpA, which mediates alveolar permeability and  H .  somni  invasion. A naturally occurring IbpA negative asymptomatic carrier strain of  H .  somni  (129Pt) does not cause BAT2 cell retraction or permeability of alveolar cell monolayers, so lacks virulence  in vitro . To investigate initial steps of pathogenesis, we showed that strain 129Pt attached to BT cells and induced a strong viperin response  in vitro . Thus colonization of the bovine upper respiratory tract with an asymptomatic carrier strain lacking virulence may decrease viral infection and the subsequent enhancement of bacterial respiratory infection  in vivo .

Histophilus somni Stimulates Expression of Antiviral Proteins and Inhibits BRSV Replication in Bovine Respiratory Epithelial Cells