Language
English
Publication Date
9-1-2023
Journal
American Journal of Veterinary Research
DOI
10.2460/ajvr.23.06.0132
PMID
37442546
Abstract
Objective: To examine the susceptibility of cultured primary equine bronchial epithelial cells (EBECs) to a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pseudovirus relative to human bronchial epithelial cells (HBECs).
Sample: Primary EBEC cultures established from healthy adult horses and commercially sourced human bronchial epithelial cells (HBECs) were used as a positive control.
Methods: Angiotensin-converting enzyme 2 (ACE2) expression by EBECs was demonstrated using immunofluorescence, western immunoblot, and flow cytometry. EBECs were transduced with a lentivirus pseudotyped with the SARS-CoV-2 spike protein that binds to ACE2 and expresses the enhanced green fluorescent protein (eGFP) as a reporter. Cells were transduced with the pseudovirus at a multiplicity of infection of 0.1 for 6 hours, washed, and maintained in media for 96 hours. After 96 hours, eGFP expression in EBECs was assessed by fluorescence microscopy of cell cultures and quantitative PCR.
Results: ACE2 expression in EBECs detected by immunofluorescence, western immunoblotting, and flow cytometry was lower in EBECs than in HBECs. After 96 hours, eGFP expression in EBECs was demonstrated by fluorescence microscopy, and mean ΔCt values from quantitative PCR were significantly (P < .0001) higher in EBECs (8.78) than HBECs (3.24) indicating lower infectivity in EBECs.
Clinical relevance: Equine respiratory tract cells were susceptible to cell entry with a SARS-CoV-2 pseudovirus. Lower replication efficiency in EBECs suggests that horses are unlikely to be an important zoonotic host of SARS-CoV-2, but viral mutations could render some strains more infective to horses. Serological and virological monitoring of horses in contact with persons shedding SARS-CoV-2 is warranted.
Keywords
Horses, Animals, Humans, SARS-CoV-2, Angiotensin-Converting Enzyme 2, Virus Internalization, COVID-19, Epithelial Cells, Horse Diseases, SARS-CoV-2, angiotensin-converting enzyme 2, horse, pseudovirus, respiratory infection
Published Open-Access
yes
Recommended Citation
Legere, Rebecca M; Allegro, Angelica R; Affram, Yvonne; et al., "Equine Bronchial Epithelial Cells Are Susceptible to Cell Entry With a SARS-CoV-2 Pseudovirus but Reveal Low Replication Efficiency" (2023). Faculty and Staff Publications. 5684.
https://digitalcommons.library.tmc.edu/baylor_docs/5684
Included in
Allergy and Immunology Commons, Biological Phenomena, Cell Phenomena, and Immunity Commons, Clinical Epidemiology Commons, COVID-19 Commons, Pathology Commons