Establishment of caprine airway epithelial cells grown in an air-liquid interface system to study caprine respiratory viruses and bacteria

Marina Strässle; Laura Laloli; Mitra Gultom; Philip V'kovski; Michael H Stoffel; Silvia Crespo Pomar; Astrid Chanfon Bätzner; Nadine Ebert; Fabien Labroussaa; Ronald Dijkman; Joerg Jores; Volker Thiel

doi:10.1016/j.vetmic.2021.109067

Establishment of caprine airway epithelial cells grown in an air-liquid interface system to study caprine respiratory viruses and bacteria

Vet Microbiol. 2021 Jun:257:109067. doi: 10.1016/j.vetmic.2021.109067. Epub 2021 Apr 10.

Authors

Affiliations

¹ Institute of Virology and Immunology (IVI), Bern, Switzerland; Institute of Veterinary Bacteriology, Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, Länggass-Str. 122, PO Box 3001, Bern, Switzerland; Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland.
² Institute of Virology and Immunology (IVI), Bern, Switzerland; Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland; Graduate School for Biomedical Science, University of Bern, Bern, Switzerland; Institute for Infectious Diseases, University of Bern, Bern, Switzerland.
³ Institute of Virology and Immunology (IVI), Bern, Switzerland; Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland.
⁴ Division of Veterinary Anatomy, Vetsuisse Faculty, University of Bern, Länggass-Str. 120, PO Box 3001, Bern, Switzerland.
⁵ Institute of Veterinary Bacteriology, Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, Länggass-Str. 122, PO Box 3001, Bern, Switzerland.
⁶ Institute of Animal Pathology (COMPATH), Vetsuisse Faculty, University of Bern, Länggass-Str. 122, PO Box 3001, Bern, Switzerland.
⁷ Institute of Virology and Immunology (IVI), Bern, Switzerland; Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland; Institute for Infectious Diseases, University of Bern, Bern, Switzerland.
⁸ Institute of Veterinary Bacteriology, Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, Länggass-Str. 122, PO Box 3001, Bern, Switzerland. Electronic address: [email protected].

PMID: 33862331
DOI: 10.1016/j.vetmic.2021.109067

Abstract

Respiratory diseases negatively impact the global goat industry, but are understudied. There is a shortage of established and biological relevant in vitro or ex vivo assays to study caprine respiratory infections. Here, we describe the establishment of an in vitro system based on well-differentiated caprine airway epithelial cell (AEC) cultures grown under air liquid interface conditions as an experimental platform to study caprine respiratory pathogens. The functional differentiation of the AEC cultures was monitored and confirmed by light and immunofluorescence microscopy, scanning electron microscopy and examination of histological sections. We validated the functionality of the platform by studying Influenza D Virus (IDV) infection and Mycoplasma mycoides subsp. capri (Mmc) colonization over 5 days, including monitoring of infectious agents by titration and qPCR as well as colour changing units, respectively. The inoculation of caprine AEC cultures with IDV showed that efficient viral replication takes place, and revealed that IDV has a marked cell tropism for ciliated cells. Furthermore, AEC cultures were successfully infected with Mmc using a multiplicity of infection of 0.1 and colonization was monitored over several days. Altogether, these results demonstrate that our newly-established caprine AEC cultures can be used to investigate host-pathogen interactions of caprine respiratory pathogens.

Keywords: AECs; ALI; Air liquid interface; Airway epithelial cell culture; Goat; Influenza D virus; Mycoplasma mycoides subsp. capri.

MeSH terms

Animals
Bronchi / cytology
Cell Culture Techniques / methods*
Cell Culture Techniques / veterinary*
Cell Differentiation
Cells, Cultured
Epithelial Cells / microbiology*
Epithelial Cells / virology*
Goats
Host-Pathogen Interactions
Microscopy, Electron, Scanning
Mycoplasma / physiology
Respiratory Mucosa / microbiology*
Respiratory Mucosa / virology*
Respiratory System / cytology*
Thogotovirus / physiology
Viral Tropism
Virus Replication / physiology

Supplementary concepts

Mycoplasma mycoides subsp. capri