Cargando…

Functional analysis of bovine TLR5 and association with IgA responses of cattle following systemic immunisation with H7 flagella

Flagellin subunits are important inducers of host immune responses through activation of TLR5 when extracellular and the inflammasome if cytosolic. Our previous work demonstrated that systemic immunization of cattle with flagella generates systemic and mucosal IgA responses. The IgA response in mice...

Descripción completa

Detalles Bibliográficos
Autores principales: Tahoun, Amin, Jensen, Kirsty, Corripio-Miyar, Yolanda, McAteer, Sean P, Corbishley, Alexander, Mahajan, Arvind, Brown, Helen, Frew, David, Aumeunier, Aude, Smith, David GE, McNeilly, Tom N, Glass, Elizabeth J, Gally, David L
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4333180/
https://www.ncbi.nlm.nih.gov/pubmed/25827709
http://dx.doi.org/10.1186/s13567-014-0135-2
_version_ 1782357989977489408
author Tahoun, Amin
Jensen, Kirsty
Corripio-Miyar, Yolanda
McAteer, Sean P
Corbishley, Alexander
Mahajan, Arvind
Brown, Helen
Frew, David
Aumeunier, Aude
Smith, David GE
McNeilly, Tom N
Glass, Elizabeth J
Gally, David L
author_facet Tahoun, Amin
Jensen, Kirsty
Corripio-Miyar, Yolanda
McAteer, Sean P
Corbishley, Alexander
Mahajan, Arvind
Brown, Helen
Frew, David
Aumeunier, Aude
Smith, David GE
McNeilly, Tom N
Glass, Elizabeth J
Gally, David L
author_sort Tahoun, Amin
collection PubMed
description Flagellin subunits are important inducers of host immune responses through activation of TLR5 when extracellular and the inflammasome if cytosolic. Our previous work demonstrated that systemic immunization of cattle with flagella generates systemic and mucosal IgA responses. The IgA response in mice is TLR5-dependent and TLR5 can impact on the general magnitude of the adaptive response. However, due to sequence differences between bovine and human/murine TLR5 sequences, it is not clear whether bovine TLR5 (bTLR5) is able to stimulate an inflammatory response following interaction with flagellin. To address this we have examined the innate responses of both human and bovine cells containing bTLR5 to H7 flagellin from E. coli O157:H7. Both HEK293 (human origin) and embryonic bovine lung (EBL) cells transfected with bTLR5 responded to addition of H7 flagellin compared to non-transfected controls. Responses were significantly reduced when mutations were introduced into the TLR5-binding regions of H7 flagellin, including an R90T substitution. In bovine primary macrophages, flagellin-stimulated CXCL8 mRNA and secreted protein levels were significantly reduced when TLR5 transcript levels were suppressed by specific siRNAs and stimulation was reduced with the R90T-H7 variant. While these results indicate that the bTLR5 sequence produces a functional flagellin-recognition receptor, cattle immunized with R90T-H7 flagella also demonstrated systemic IgA responses to the flagellin in comparison to adjuvant only controls. This presumably either reflects our findings that R90T-H7 still activates bTLR5, albeit with reduced efficiency compared to WT H7 flagellin, or that other flagellin recognition pathways may play a role in this mucosal response.
format Online
Article
Text
id pubmed-4333180
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-43331802015-02-20 Functional analysis of bovine TLR5 and association with IgA responses of cattle following systemic immunisation with H7 flagella Tahoun, Amin Jensen, Kirsty Corripio-Miyar, Yolanda McAteer, Sean P Corbishley, Alexander Mahajan, Arvind Brown, Helen Frew, David Aumeunier, Aude Smith, David GE McNeilly, Tom N Glass, Elizabeth J Gally, David L Vet Res Research Flagellin subunits are important inducers of host immune responses through activation of TLR5 when extracellular and the inflammasome if cytosolic. Our previous work demonstrated that systemic immunization of cattle with flagella generates systemic and mucosal IgA responses. The IgA response in mice is TLR5-dependent and TLR5 can impact on the general magnitude of the adaptive response. However, due to sequence differences between bovine and human/murine TLR5 sequences, it is not clear whether bovine TLR5 (bTLR5) is able to stimulate an inflammatory response following interaction with flagellin. To address this we have examined the innate responses of both human and bovine cells containing bTLR5 to H7 flagellin from E. coli O157:H7. Both HEK293 (human origin) and embryonic bovine lung (EBL) cells transfected with bTLR5 responded to addition of H7 flagellin compared to non-transfected controls. Responses were significantly reduced when mutations were introduced into the TLR5-binding regions of H7 flagellin, including an R90T substitution. In bovine primary macrophages, flagellin-stimulated CXCL8 mRNA and secreted protein levels were significantly reduced when TLR5 transcript levels were suppressed by specific siRNAs and stimulation was reduced with the R90T-H7 variant. While these results indicate that the bTLR5 sequence produces a functional flagellin-recognition receptor, cattle immunized with R90T-H7 flagella also demonstrated systemic IgA responses to the flagellin in comparison to adjuvant only controls. This presumably either reflects our findings that R90T-H7 still activates bTLR5, albeit with reduced efficiency compared to WT H7 flagellin, or that other flagellin recognition pathways may play a role in this mucosal response. BioMed Central 2015-02-19 2015 /pmc/articles/PMC4333180/ /pubmed/25827709 http://dx.doi.org/10.1186/s13567-014-0135-2 Text en © Tahoun et al.; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Tahoun, Amin
Jensen, Kirsty
Corripio-Miyar, Yolanda
McAteer, Sean P
Corbishley, Alexander
Mahajan, Arvind
Brown, Helen
Frew, David
Aumeunier, Aude
Smith, David GE
McNeilly, Tom N
Glass, Elizabeth J
Gally, David L
Functional analysis of bovine TLR5 and association with IgA responses of cattle following systemic immunisation with H7 flagella
title Functional analysis of bovine TLR5 and association with IgA responses of cattle following systemic immunisation with H7 flagella
title_full Functional analysis of bovine TLR5 and association with IgA responses of cattle following systemic immunisation with H7 flagella
title_fullStr Functional analysis of bovine TLR5 and association with IgA responses of cattle following systemic immunisation with H7 flagella
title_full_unstemmed Functional analysis of bovine TLR5 and association with IgA responses of cattle following systemic immunisation with H7 flagella
title_short Functional analysis of bovine TLR5 and association with IgA responses of cattle following systemic immunisation with H7 flagella
title_sort functional analysis of bovine tlr5 and association with iga responses of cattle following systemic immunisation with h7 flagella
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4333180/
https://www.ncbi.nlm.nih.gov/pubmed/25827709
http://dx.doi.org/10.1186/s13567-014-0135-2
work_keys_str_mv AT tahounamin functionalanalysisofbovinetlr5andassociationwithigaresponsesofcattlefollowingsystemicimmunisationwithh7flagella
AT jensenkirsty functionalanalysisofbovinetlr5andassociationwithigaresponsesofcattlefollowingsystemicimmunisationwithh7flagella
AT corripiomiyaryolanda functionalanalysisofbovinetlr5andassociationwithigaresponsesofcattlefollowingsystemicimmunisationwithh7flagella
AT mcateerseanp functionalanalysisofbovinetlr5andassociationwithigaresponsesofcattlefollowingsystemicimmunisationwithh7flagella
AT corbishleyalexander functionalanalysisofbovinetlr5andassociationwithigaresponsesofcattlefollowingsystemicimmunisationwithh7flagella
AT mahajanarvind functionalanalysisofbovinetlr5andassociationwithigaresponsesofcattlefollowingsystemicimmunisationwithh7flagella
AT brownhelen functionalanalysisofbovinetlr5andassociationwithigaresponsesofcattlefollowingsystemicimmunisationwithh7flagella
AT frewdavid functionalanalysisofbovinetlr5andassociationwithigaresponsesofcattlefollowingsystemicimmunisationwithh7flagella
AT aumeunieraude functionalanalysisofbovinetlr5andassociationwithigaresponsesofcattlefollowingsystemicimmunisationwithh7flagella
AT smithdavidge functionalanalysisofbovinetlr5andassociationwithigaresponsesofcattlefollowingsystemicimmunisationwithh7flagella
AT mcneillytomn functionalanalysisofbovinetlr5andassociationwithigaresponsesofcattlefollowingsystemicimmunisationwithh7flagella
AT glasselizabethj functionalanalysisofbovinetlr5andassociationwithigaresponsesofcattlefollowingsystemicimmunisationwithh7flagella
AT gallydavidl functionalanalysisofbovinetlr5andassociationwithigaresponsesofcattlefollowingsystemicimmunisationwithh7flagella