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Sheep primary cells as in vitro models to investigate Mycoplasma agalactiae host cell interactions

Appropriate infection models are imperative for the understanding of pathogens like mycoplasmas that are known for their strict host and tissue specificity, and lack of suitable cell and small animal models has hindered pathogenicity studies. This is particularly true for the economically important...

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Autores principales: Hegde, Shrilakshmi, Gabriel, Cordula, Kragl, Martin, Chopra-Dewasthaly, Rohini
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4535462/
https://www.ncbi.nlm.nih.gov/pubmed/26187893
http://dx.doi.org/10.1093/femspd/ftv048
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author Hegde, Shrilakshmi
Gabriel, Cordula
Kragl, Martin
Chopra-Dewasthaly, Rohini
author_facet Hegde, Shrilakshmi
Gabriel, Cordula
Kragl, Martin
Chopra-Dewasthaly, Rohini
author_sort Hegde, Shrilakshmi
collection PubMed
description Appropriate infection models are imperative for the understanding of pathogens like mycoplasmas that are known for their strict host and tissue specificity, and lack of suitable cell and small animal models has hindered pathogenicity studies. This is particularly true for the economically important group of ruminant mycoplasmas whose virulence factors need to be elucidated for designing effective intervention strategies. Mycoplasma agalactiae serves as a useful role model especially because it is phylogenetically very close to M. bovis and causes similar symptoms by as yet unknown mechanisms. Here, we successfully prepared and characterized four different primary sheep cell lines, namely the epithelial and stromal cells from the mammary gland and uterus, respectively. Using immunohistochemistry, we identified vimentin and cytokeratin as specific markers to confirm the typical cell phenotypes of these primary cells. Furthermore, M. agalactiae’s consistent adhesion and invasion into these primary cells proves the reliability of these cell models. Mimicking natural infections, mammary epithelial and stromal cells showed higher invasion and adhesion rates compared to the uterine cells as also seen via double immunofluorescence staining. Altogether, we have generated promising in vitro cell models to study host–pathogen interactions of M. agalactiae and related ruminant pathogens in a more authentic manner.
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spelling pubmed-45354622015-08-17 Sheep primary cells as in vitro models to investigate Mycoplasma agalactiae host cell interactions Hegde, Shrilakshmi Gabriel, Cordula Kragl, Martin Chopra-Dewasthaly, Rohini Pathog Dis Short Communication Appropriate infection models are imperative for the understanding of pathogens like mycoplasmas that are known for their strict host and tissue specificity, and lack of suitable cell and small animal models has hindered pathogenicity studies. This is particularly true for the economically important group of ruminant mycoplasmas whose virulence factors need to be elucidated for designing effective intervention strategies. Mycoplasma agalactiae serves as a useful role model especially because it is phylogenetically very close to M. bovis and causes similar symptoms by as yet unknown mechanisms. Here, we successfully prepared and characterized four different primary sheep cell lines, namely the epithelial and stromal cells from the mammary gland and uterus, respectively. Using immunohistochemistry, we identified vimentin and cytokeratin as specific markers to confirm the typical cell phenotypes of these primary cells. Furthermore, M. agalactiae’s consistent adhesion and invasion into these primary cells proves the reliability of these cell models. Mimicking natural infections, mammary epithelial and stromal cells showed higher invasion and adhesion rates compared to the uterine cells as also seen via double immunofluorescence staining. Altogether, we have generated promising in vitro cell models to study host–pathogen interactions of M. agalactiae and related ruminant pathogens in a more authentic manner. Oxford University Press 2015-07-17 2015-10 /pmc/articles/PMC4535462/ /pubmed/26187893 http://dx.doi.org/10.1093/femspd/ftv048 Text en © FEMS 2015. http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits non-commercial reuse, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Short Communication
Hegde, Shrilakshmi
Gabriel, Cordula
Kragl, Martin
Chopra-Dewasthaly, Rohini
Sheep primary cells as in vitro models to investigate Mycoplasma agalactiae host cell interactions
title Sheep primary cells as in vitro models to investigate Mycoplasma agalactiae host cell interactions
title_full Sheep primary cells as in vitro models to investigate Mycoplasma agalactiae host cell interactions
title_fullStr Sheep primary cells as in vitro models to investigate Mycoplasma agalactiae host cell interactions
title_full_unstemmed Sheep primary cells as in vitro models to investigate Mycoplasma agalactiae host cell interactions
title_short Sheep primary cells as in vitro models to investigate Mycoplasma agalactiae host cell interactions
title_sort sheep primary cells as in vitro models to investigate mycoplasma agalactiae host cell interactions
topic Short Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4535462/
https://www.ncbi.nlm.nih.gov/pubmed/26187893
http://dx.doi.org/10.1093/femspd/ftv048
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