Cargando…

Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent

Bovine mastitis is a costly disease to the dairy industry and intramammary infections (IMI) with Staphylococcus aureus are a major cause of mastitis. Staphylococcus aureus strains responsible for mastitis in cattle predominantly belong to ruminant-associated clonal complexes (CCs). Recognition of pa...

Descripción completa

Detalles Bibliográficos
Autores principales: Hoekstra, Jurriaan, Rutten, Victor P. M. G., Lam, Theo J. G. M., Van Kessel, Kok P. M., Spaninks, Mirlin P., Stegeman, J. Arjan, Benedictus, Lindert, Koop, Gerrit
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6955728/
https://www.ncbi.nlm.nih.gov/pubmed/31842337
http://dx.doi.org/10.3390/microorganisms7120688
_version_ 1783486995036635136
author Hoekstra, Jurriaan
Rutten, Victor P. M. G.
Lam, Theo J. G. M.
Van Kessel, Kok P. M.
Spaninks, Mirlin P.
Stegeman, J. Arjan
Benedictus, Lindert
Koop, Gerrit
author_facet Hoekstra, Jurriaan
Rutten, Victor P. M. G.
Lam, Theo J. G. M.
Van Kessel, Kok P. M.
Spaninks, Mirlin P.
Stegeman, J. Arjan
Benedictus, Lindert
Koop, Gerrit
author_sort Hoekstra, Jurriaan
collection PubMed
description Bovine mastitis is a costly disease to the dairy industry and intramammary infections (IMI) with Staphylococcus aureus are a major cause of mastitis. Staphylococcus aureus strains responsible for mastitis in cattle predominantly belong to ruminant-associated clonal complexes (CCs). Recognition of pathogens by bovine mammary epithelial cells (bMEC) plays a key role in activation of immune responsiveness during IMI. However, it is still largely unknown to what extent the bMEC response differs according to S. aureus CC. The aim of this study was to determine whether ruminant-associated S. aureus CCs differentially activate bMEC. For this purpose, the immortalized bMEC line PS was stimulated with S. aureus mastitis isolates belonging to four different clonal complexes (CCs; CC133, CC479, CC151 and CC425) and interleukin 8 (IL-8) release was measured as indicator of activation. To validate our bMEC model, we first stimulated PS cells with genetically modified S. aureus strains lacking (protein A, wall teichoic acid (WTA) synthesis) or expressing (capsular polysaccharide (CP) type 5 or type 8) factors expected to affect S. aureus recognition by bMEC. The absence of functional WTA synthesis increased IL-8 release by bMEC in response to bacterial stimulation compared to wildtype. In addition, bMEC released more IL-8 after stimulation with S. aureus expressing CP type 5 compared to CP type 8 or a strain lacking CP expression. Among the S. aureus lineages, isolates belonging to CC133 induced a significantly stronger IL-8 release from bMEC than isolates from the other CCs, and the IL-8 response to CC479 was higher compared to CC151 and CC425. Transcription levels of IL-8, tumor necrosis factor alpha (TNFα), serum amyloid A3 (SAA3), Toll-like receptor (TLR)-2 and nuclear factor κB (NF-κB) in bMEC after bacterial stimulation tended to follow a similar pattern as IL-8 release, but there were no significant differences between the CCs. This study demonstrates a differential activation of bMEC by ruminant-associated CCs of S. aureus, which may have implications for the severity of mastitis during IMI by S. aureus belonging to these lineages.
format Online
Article
Text
id pubmed-6955728
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-69557282020-01-23 Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent Hoekstra, Jurriaan Rutten, Victor P. M. G. Lam, Theo J. G. M. Van Kessel, Kok P. M. Spaninks, Mirlin P. Stegeman, J. Arjan Benedictus, Lindert Koop, Gerrit Microorganisms Article Bovine mastitis is a costly disease to the dairy industry and intramammary infections (IMI) with Staphylococcus aureus are a major cause of mastitis. Staphylococcus aureus strains responsible for mastitis in cattle predominantly belong to ruminant-associated clonal complexes (CCs). Recognition of pathogens by bovine mammary epithelial cells (bMEC) plays a key role in activation of immune responsiveness during IMI. However, it is still largely unknown to what extent the bMEC response differs according to S. aureus CC. The aim of this study was to determine whether ruminant-associated S. aureus CCs differentially activate bMEC. For this purpose, the immortalized bMEC line PS was stimulated with S. aureus mastitis isolates belonging to four different clonal complexes (CCs; CC133, CC479, CC151 and CC425) and interleukin 8 (IL-8) release was measured as indicator of activation. To validate our bMEC model, we first stimulated PS cells with genetically modified S. aureus strains lacking (protein A, wall teichoic acid (WTA) synthesis) or expressing (capsular polysaccharide (CP) type 5 or type 8) factors expected to affect S. aureus recognition by bMEC. The absence of functional WTA synthesis increased IL-8 release by bMEC in response to bacterial stimulation compared to wildtype. In addition, bMEC released more IL-8 after stimulation with S. aureus expressing CP type 5 compared to CP type 8 or a strain lacking CP expression. Among the S. aureus lineages, isolates belonging to CC133 induced a significantly stronger IL-8 release from bMEC than isolates from the other CCs, and the IL-8 response to CC479 was higher compared to CC151 and CC425. Transcription levels of IL-8, tumor necrosis factor alpha (TNFα), serum amyloid A3 (SAA3), Toll-like receptor (TLR)-2 and nuclear factor κB (NF-κB) in bMEC after bacterial stimulation tended to follow a similar pattern as IL-8 release, but there were no significant differences between the CCs. This study demonstrates a differential activation of bMEC by ruminant-associated CCs of S. aureus, which may have implications for the severity of mastitis during IMI by S. aureus belonging to these lineages. MDPI 2019-12-12 /pmc/articles/PMC6955728/ /pubmed/31842337 http://dx.doi.org/10.3390/microorganisms7120688 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Hoekstra, Jurriaan
Rutten, Victor P. M. G.
Lam, Theo J. G. M.
Van Kessel, Kok P. M.
Spaninks, Mirlin P.
Stegeman, J. Arjan
Benedictus, Lindert
Koop, Gerrit
Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent
title Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent
title_full Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent
title_fullStr Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent
title_full_unstemmed Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent
title_short Activation of a Bovine Mammary Epithelial Cell Line by Ruminant-Associated Staphylococcus aureus is Lineage Dependent
title_sort activation of a bovine mammary epithelial cell line by ruminant-associated staphylococcus aureus is lineage dependent
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6955728/
https://www.ncbi.nlm.nih.gov/pubmed/31842337
http://dx.doi.org/10.3390/microorganisms7120688
work_keys_str_mv AT hoekstrajurriaan activationofabovinemammaryepithelialcelllinebyruminantassociatedstaphylococcusaureusislineagedependent
AT ruttenvictorpmg activationofabovinemammaryepithelialcelllinebyruminantassociatedstaphylococcusaureusislineagedependent
AT lamtheojgm activationofabovinemammaryepithelialcelllinebyruminantassociatedstaphylococcusaureusislineagedependent
AT vankesselkokpm activationofabovinemammaryepithelialcelllinebyruminantassociatedstaphylococcusaureusislineagedependent
AT spaninksmirlinp activationofabovinemammaryepithelialcelllinebyruminantassociatedstaphylococcusaureusislineagedependent
AT stegemanjarjan activationofabovinemammaryepithelialcelllinebyruminantassociatedstaphylococcusaureusislineagedependent
AT benedictuslindert activationofabovinemammaryepithelialcelllinebyruminantassociatedstaphylococcusaureusislineagedependent
AT koopgerrit activationofabovinemammaryepithelialcelllinebyruminantassociatedstaphylococcusaureusislineagedependent