Development of a primary cell model derived from porcine dorsal soft palate for foot-and-mouth disease virus research and diagnosis - Anses - Agence nationale de sécurité sanitaire de l’alimentation, de l’environnement et du travail Accéder directement au contenu
Poster De Conférence Année : 2023

Development of a primary cell model derived from porcine dorsal soft palate for foot-and-mouth disease virus research and diagnosis

Morgan Sarry
Anthony Relmy
  • Fonction : Auteur
  • PersonId : 1075108
Aurore Romey
Hélène Huet

Résumé

Background and Objectives Foot-and-mouth disease (FMD) is a contagious and devastating animal disease affecting artiodactyls. The persistence of FMDV in some ruminants after clinical recovery remains problematic. Such persistence is defined as the presence of infectious virus in the host beyond 28 days post-infection (dpi). Persistent FMDV has been found in dorsal soft palate (DSP) epithelial cells in ruminants. While the DSP is one of the primary sites of virus replication in both ruminants and swine, no persistent infectious virus was found in swine DSP. Only viral RNA could be detected in vivo over 60 dpi. In order to improve FMDV persistence knowledge, we have developed an in vitro model to overcome the lack of porcine suitable models. Material and Methods Primary epithelial cells were isolated from DSP tissues collected from 4-6 month old swine. Both the presence of epithelial markers in these cells, such as occludin, and the presence of FMDV-specific integrin receptors were assessed by immunofluorescence. Molecular analyses were performed to check that the cells were free of viruses commonly found in swine. Once characterised, the cells were inoculated as monolayers and as multilayers cultivated at the air-liquid interface (ALI), with a type-O FMDV. Samples were regularly collected up to 115 dpi and were tested for the presence of infectious virus, viral RNA and viral antigen. Results We confirmed that these cells were epithelial, possessed FMDV-specific receptors and were free of three common porcine viruses. Experimental infections revealed that they were less sensitive to FMDV than the bovine DSP model we previously established. Analysis of collected supernatants identified infectious virus up to 14 dpi, as well as viral RNA up to 60 dpi, consistent with in vivo observations. Multilayer (ALI) cell culture mimicked a multilayered epithelium more similar to the biological system. Although viral RNA were detected up to 35 dpi, no infectious viruses were however detected in the supernatants after infection of the multilayers. Conclusion The development and infection of the DSP primary porcine cells confirmed the absence of any evidence of FMDV persistence in this swine cell model as no infectious virus could be detected after 28 days. The relevance of this cellular model having been confirmed by the results obtained, it would be considered to immortalise these cells in view of developing a porcine epithelial cell line that could be used for research and FMD diagnosis.
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Dates et versions

anses-04426176 , version 1 (30-01-2024)

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Paternité - Pas d'utilisation commerciale - Pas de modification

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  • HAL Id : anses-04426176 , version 1

Citer

Morgan Sarry, Cindy Bernelin Cottet, Caroline Michaud, Anthony Relmy, Aurore Romey, et al.. Development of a primary cell model derived from porcine dorsal soft palate for foot-and-mouth disease virus research and diagnosis. Epizone, 15th annual meeting 2023, Apr 2023, Novi Sad, Serbia. ⟨anses-04426176⟩
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