Gene signatures associated with foot-and-mouth disease virus infection and persistence part I: persistent FMDV in a three-dimensional model of the bovine soft palate
Résumé
Foot-and-mouth disease virus (FMDV) causes a highly contagious vesicular disease in livestock, with serious consequences for international trade. FMDV persistence in the oropharynx of cattle slows down the process to obtain an FMDV-free status after an outbreak. To study biological mechanisms, or to identify molecules that can be targeted to diagnose or interfere with persistence, we developed a model of persistent FMDV infection in bovine dorsal soft palate (DSP). Primary DSP cells were cultured in multilayers at the air-liquid interface (ALI). After 5 weeks of culture without further passage, the cells were infected with FMDV strain O/FRA/1/2001. Infection was monitored until 28 DPI. Approximately 20% of cells still had a polygonal morphology and displayed tight junctions as in stratified squamous epithelia. Cells with similar morphology expressed cytokeratin. A limited cytopathic effect was induced, restricted to the upper cell layers. FMDV antigen, FMDV RNA and live virus were detected from day 1 to 28, with peaks at day 1 and 2. The proportion of infected cells was highest at 24 hours (3 % and 36 % of cells at an MOI of 0.01 and 1, respectively). At day 28 after infection, at a time when animals that still harbour FMDV are considered carriers, FMDV antigen was detected in 0.2% - 2.1% of cells, in all layers, and live virus was isolated from supernatants of 6/8 cultures. The ALI model of DSP brings new possibilities to investigate FMDV persistence in a controlled manner. Transcriptomic data will be presented in a joint communication.
Domaines
Sciences du Vivant [q-bio]Origine | Fichiers produits par l'(les) auteur(s) |
---|