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

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.

Host Range, Biology, and Species Specificity of Seven-Segmented Influenza Viruses—A Comparative Review on Influenza C and D

“…Compared with ICV, IDV has broad cell tropism owing to its increased binding avidity towards diverse glycans and can grow not only in continuous cell lines, but also in most of the primary cells from bovine [162], caprine [163], equine, and porcine species [164]. A comparison of in vitro cellular tropism of the IDV to ICV, determined by RT-PCR in cell lines of different species origins, such as swine testicle (ST) cells [165], (MDCK), green African monkey kidney cells (Marc-145), human rectal tumor (HRT-18G), baby hamster kidney (BHK-21), and porcine kidney (PK-15) cells, showed that IDV replicated to appreciable titers in all, with minimal replication in BHK-21 and PK-15 cells [82].…”

Galleria mellonella Invertebrate Model Mirrors the Pathogenic Potential of Mycoplasma alligatoris within the Natural Host

“…Current methods to determine virulence potential and host-pathogen interactions of mycoplasmas have relied on (i) in vivo experimental infections of the natural or alternative hosts [32][33][34][35] or on (ii) in vitro infections [21,[36][37][38][39][40]. Infections of the natural host, especially for reptiles, can be challenging but have been successfully used to confirm that both M. alligatoris [27,41] and M. agassizii [42,43] are etiologic agents of disease in alligators and tortoises, respectively.…”

“…In vitro assays have focused on the use of cell lines to screen a large number of Mycoplasma spp. for pathogenic potential and to understand cellular and molecular interactions [10,21,36,37,39,40,44]. However, cell lines lack a functioning immune system which is a key component of mycoplasmal disease [1,2,[45][46][47][48].…”