Equine bronchial epithelial cells are susceptible to cell entry with a SARS-CoV-2 pseudovirus but reveal low replication efficiency.
Abstract: 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). Methods: 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. Conclusions: 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.
Publication Date: 2023-07-18 PubMed ID: 37442546DOI: 10.2460/ajvr.23.06.0132Google Scholar: Lookup
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- Journal Article
Summary
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This study investigated how cells in the horse respiratory system react to a simulated SARS-CoV-2 virus, discovering that while these cells can be infected, the virus doesn’t replicate well. This suggests that horses may not be a significant host for the virus, although virus mutations could change this.
Methodology
- The study used primary cultures of equine bronchial epithelial cells (EBECs), taken from healthy adult horses.
- Human bronchial epithelial cells (HBECs) were used as a positive control group, to compare the reaction of the equine cells to a known response.
- The EBECs were tested for expression of the angiotensin-converting enzyme 2 (ACE2), the protein to which SARS-CoV-2 binds, using multiple techniques: immunofluorescence, western immunoblot, and flow cytometry.
- The EBECs were then exposed to a lentivirus pseudotyped with the SARS-CoV-2 spike protein, which shows up as enhanced green fluorescent protein (eGFP), at a rate of infection of 0.1 for six hours.
- The cells were then washed, and maintained in media for the next 96 hours.
Results
- ACE2 expression in EBECs was lower than in HBECs, as demonstrated by the three testing techniques.
- After the 96-hour period, eGFP expression was visible in the EBECs, as detected by fluorescence microscopy.
- The results of quantitative PCR showed higher ΔCt values in the EBECs (8.78) compared to the HBECs (3.24).
- The higher ΔCt values in EBECs suggest lower infectivity in these cells.
Conclusion
- Respiratory tract cells in horses can be infected by a SARS-CoV-2 pseudovirus.
- However, the efficiency of the virus replication in EBECs is low, suggesting that horses may not be a significant source of the virus in a zoonotic context.
- There is a risk that viral mutations may make some strains of SARS-CoV-2 more infectious to horses. Thus, monitoring horses in contact with humans who are shedding SARS-CoV-2 via serological and virological tests is recommended.
Cite This Article
APA
Legere RM, Allegro AR, Affram Y, Silveira BPD, Fridley JL, Wells KM, Oezguen N, Burghardt RC, Wright GA, Pollet J, Bordin AI, Figueiredo P, Leibowitz JL, Cohen ND.
(2023).
Equine bronchial epithelial cells are susceptible to cell entry with a SARS-CoV-2 pseudovirus but reveal low replication efficiency.
Am J Vet Res, 1-11.
https://doi.org/10.2460/ajvr.23.06.0132 Publication
Researcher Affiliations
- Equine Infectious Disease Laboratory, Department of Large Animal Clinical Sciences, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX.
- Equine Infectious Disease Laboratory, Department of Large Animal Clinical Sciences, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX.
- Department of Microbial Pathogenesis & Immunology, School of Medicine, Texas A&M University, College Station, TX.
- Equine Infectious Disease Laboratory, Department of Large Animal Clinical Sciences, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX.
- Department of Large Animal Clinical Sciences, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX.
- Department of Microbial Pathogenesis & Immunology, School of Medicine, Texas A&M University, College Station, TX.
- Texas Children's Microbiome Center, Baylor College of Medicine, Houston, TX.
- Department of Veterinary Integrative Biosciences, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX.
- Department of Veterinary Pathobiology, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX.
- National School of Tropical Medicine, Department of Pediatrics, Baylor College of Medicine, Houston, TX.
- Texas Children's Hospital Center for Vaccine Development, Baylor College of Medicine, Houston, TX.
- Equine Infectious Disease Laboratory, Department of Large Animal Clinical Sciences, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX.
- Department of Microbial Pathogenesis & Immunology, School of Medicine, Texas A&M University, College Station, TX.
- Department of Veterinary Pathobiology, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX.
- Department of Microbial Pathogenesis & Immunology, School of Medicine, Texas A&M University, College Station, TX.
- Equine Infectious Disease Laboratory, Department of Large Animal Clinical Sciences, School of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX.
Citations
This article has been cited 1 times.- Pucca MB, Camphora AL. The potential risks of equine serum therapy in transmitting new infectious diseases: lessons from a post-pandemic era. Front Public Health 2024;12:1366929.
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