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  1. Article ; Online: Equine bronchial epithelial cells are susceptible to cell entry with a SARS-CoV-2 pseudovirus but reveal low replication efficiency.

    Legere, Rebecca M / Allegro, Angelica R / Affram, Yvonne / Silveira, Bibiana Petri da / Fridley, Jennifer L / Wells, Kelsey M / Oezguen, Numan / Burghardt, Robert C / Wright, Gus A / Pollet, Jeroen / Bordin, Angela I / Figueiredo, Paul de / Leibowitz, Julian L / Cohen, Noah D

    American journal of veterinary research

    2023  Volume 84, Issue 9

    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: ... ...

    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.
    MeSH term(s) Horses ; Animals ; Humans ; SARS-CoV-2/metabolism ; Angiotensin-Converting Enzyme 2/metabolism ; Virus Internalization ; COVID-19/veterinary ; Epithelial Cells ; Horse Diseases
    Chemical Substances spike protein, SARS-CoV-2 ; Angiotensin-Converting Enzyme 2 (EC 3.4.17.23)
    Language English
    Publishing date 2023-07-18
    Publishing country United States
    Document type Journal Article
    ZDB-ID 390796-x
    ISSN 1943-5681 ; 0002-9645
    ISSN (online) 1943-5681
    ISSN 0002-9645
    DOI 10.2460/ajvr.23.06.0132
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Intramuscular but not nebulized administration of a mRNA vaccine against Rhodococcus equi stimulated humoral immune responses in neonatal foals.

    Legere, Rebecca M / Poveda, Cristina / Ott, Jeannine A / Bray, Jocelyne M / Villafone, Emma G / Silveira, Bibiana Petri da / Kahn, Susanne K / Martin, Cameron L / Mancino, Chiara / Taraballi, Francesca / Criscitiello, Michael F / Berghman, Luc / Bordin, Angela I / Pollet, Jeroen / Cohen, Noah D

    American journal of veterinary research

    2023  Volume 85, Issue 2

    Abstract: Objective: Design and evaluate immune responses of neonatal foals to a mRNA vaccine expressing the virulence-associated protein A (VapA) of Rhodococcus equi.: Animals: Cultured primary equine respiratory tract cells; Serum, bronchoalveolar lavage ... ...

    Abstract Objective: Design and evaluate immune responses of neonatal foals to a mRNA vaccine expressing the virulence-associated protein A (VapA) of Rhodococcus equi.
    Animals: Cultured primary equine respiratory tract cells; Serum, bronchoalveolar lavage fluid (BALF), and peripheral blood mononuclear cells (PBMCs) from 30 healthy Quarter Horse foals.
    Methods: VapA expression was evaluated by western immunoblot in cultured equine bronchial cells transfected with 4 mRNA constructs encoding VapA. The mRNA construct with greatest expression was used to immunize foals at ages 2 and 21 days in 5 groups: (1) 300 μg nebulized mRNA (n = 6); (2) 600 μg nebulized mRNA (n = 4); (3) 300 μg mRNA administered intramuscularly (IM) (n = 5); (4) 300 μg VapA IM (positive controls; n = 6); or (5) nebulized water (negative controls; n = 6). Serum, BALF, and PBMCs were collected at ages 3, 22, and 35 days and tested for relative anti-VapA IgG1, IgG4/7, and IgA activities using ELISA and cell-mediated immunity by ELISpot.
    Results: As formulated, nebulized mRNA was not immunogenic. However, a significant increase in anti-VapA IgG4/7 activity (P < .05) was noted exclusively in foals immunized IM with VapA mRNA by age 35 days. The proportion of foals with anti-VapA IgG1 activity > 30% of positive control differed significantly (P = .0441) between negative controls (50%; 3/6), IM mRNA foals (100%; 5/5), and IM VapA (100%; 6/6) groups. Natural exposure to virulent R equi was immunogenic in some negative control foals.
    Clinical relevance: Further evaluation of the immunogenicity and efficacy of IM mRNA encoding VapA in foals is warranted.
    MeSH term(s) Animals ; Horses ; Animals, Newborn ; Immunity, Humoral ; mRNA Vaccines ; Bacterial Proteins/genetics ; Rhodococcus equi/genetics ; Leukocytes, Mononuclear ; Immunoglobulin G ; RNA, Messenger/genetics ; Actinomycetales Infections/prevention & control ; Actinomycetales Infections/veterinary ; Horse Diseases/prevention & control ; Virulence Factors/genetics
    Chemical Substances mRNA Vaccines ; Bacterial Proteins ; Immunoglobulin G ; RNA, Messenger ; Virulence Factors
    Language English
    Publishing date 2023-12-11
    Publishing country United States
    Document type Journal Article
    ZDB-ID 390796-x
    ISSN 1943-5681 ; 0002-9645
    ISSN (online) 1943-5681
    ISSN 0002-9645
    DOI 10.2460/ajvr.23.09.0208
    Database MEDical Literature Analysis and Retrieval System OnLINE

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