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Respiratory explants as a model to investigate early events of contagious bovine pleuropneumonia infection
Contagious bovine pleuropneumonia (CBPP) is a severe disease caused by Mycoplasma mycoides subsp. mycoides (Mmm). Knowledge on CBPP pathogenesis is fragmented and hampered by the limited availability of laboratory animal and in vitro models of investigation. The purpose of the present study is to as...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5766988/ https://www.ncbi.nlm.nih.gov/pubmed/29329577 http://dx.doi.org/10.1186/s13567-017-0500-z |
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author | Di Teodoro, Giovanni Marruchella, Giuseppe Di Provvido, Andrea Orsini, Gianluca Ronchi, Gaetano Federico D’Angelo, Anna Rita D’Alterio, Nicola Sacchini, Flavio Scacchia, Massimo |
author_facet | Di Teodoro, Giovanni Marruchella, Giuseppe Di Provvido, Andrea Orsini, Gianluca Ronchi, Gaetano Federico D’Angelo, Anna Rita D’Alterio, Nicola Sacchini, Flavio Scacchia, Massimo |
author_sort | Di Teodoro, Giovanni |
collection | PubMed |
description | Contagious bovine pleuropneumonia (CBPP) is a severe disease caused by Mycoplasma mycoides subsp. mycoides (Mmm). Knowledge on CBPP pathogenesis is fragmented and hampered by the limited availability of laboratory animal and in vitro models of investigation. The purpose of the present study is to assess respiratory explants as useful tools to study the early stages of CBPP. Explants were obtained from trachea, bronchi and lungs of slaughtered cattle, tested negative for Mycoplasma spp. and for the major bacterial and viral respiratory pathogens. The interaction of Mmm with explant cells was studied by immunohistochemistry (IHC), double-labelling indirect immunofluorescence (DLIIF) and laser scanning confocal microscopy (LSCM). Mmm capability to survive and proliferate within the explants was evaluated by standard microbiological procedures. Finally, the putative cellular internalization of Mmm was further investigated by the gentamicin invasion assay. IHC and DLIIF indicated that Mmm can colonize explants, showing a marked tropism for lower airways. Specifically, Mmm was detected on/inside the bronchiolar and alveolar epithelial cells, the alveolar macrophages and the endothelial cells. The interaction between Mmm and explant cells was abolished by the pre-incubation of the pathogen with bovine anti-Mmm immune sera. Mmm was able to survive and proliferate in all tracheal, bronchial and lung explants, during the entire time course of the experiments. LSCM and gentamicin invasion assay both confirmed that Mmm can enter non-phagocytic host cells. Taken together, our data supports bovine respiratory explants as a promising tool to investigate CBPP, alternative to cattle experimental infection. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13567-017-0500-z) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5766988 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-57669882018-01-17 Respiratory explants as a model to investigate early events of contagious bovine pleuropneumonia infection Di Teodoro, Giovanni Marruchella, Giuseppe Di Provvido, Andrea Orsini, Gianluca Ronchi, Gaetano Federico D’Angelo, Anna Rita D’Alterio, Nicola Sacchini, Flavio Scacchia, Massimo Vet Res Research Article Contagious bovine pleuropneumonia (CBPP) is a severe disease caused by Mycoplasma mycoides subsp. mycoides (Mmm). Knowledge on CBPP pathogenesis is fragmented and hampered by the limited availability of laboratory animal and in vitro models of investigation. The purpose of the present study is to assess respiratory explants as useful tools to study the early stages of CBPP. Explants were obtained from trachea, bronchi and lungs of slaughtered cattle, tested negative for Mycoplasma spp. and for the major bacterial and viral respiratory pathogens. The interaction of Mmm with explant cells was studied by immunohistochemistry (IHC), double-labelling indirect immunofluorescence (DLIIF) and laser scanning confocal microscopy (LSCM). Mmm capability to survive and proliferate within the explants was evaluated by standard microbiological procedures. Finally, the putative cellular internalization of Mmm was further investigated by the gentamicin invasion assay. IHC and DLIIF indicated that Mmm can colonize explants, showing a marked tropism for lower airways. Specifically, Mmm was detected on/inside the bronchiolar and alveolar epithelial cells, the alveolar macrophages and the endothelial cells. The interaction between Mmm and explant cells was abolished by the pre-incubation of the pathogen with bovine anti-Mmm immune sera. Mmm was able to survive and proliferate in all tracheal, bronchial and lung explants, during the entire time course of the experiments. LSCM and gentamicin invasion assay both confirmed that Mmm can enter non-phagocytic host cells. Taken together, our data supports bovine respiratory explants as a promising tool to investigate CBPP, alternative to cattle experimental infection. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13567-017-0500-z) contains supplementary material, which is available to authorized users. BioMed Central 2018-01-12 2018 /pmc/articles/PMC5766988/ /pubmed/29329577 http://dx.doi.org/10.1186/s13567-017-0500-z Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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 Article Di Teodoro, Giovanni Marruchella, Giuseppe Di Provvido, Andrea Orsini, Gianluca Ronchi, Gaetano Federico D’Angelo, Anna Rita D’Alterio, Nicola Sacchini, Flavio Scacchia, Massimo Respiratory explants as a model to investigate early events of contagious bovine pleuropneumonia infection |
title | Respiratory explants as a model to investigate early events of contagious bovine pleuropneumonia infection |
title_full | Respiratory explants as a model to investigate early events of contagious bovine pleuropneumonia infection |
title_fullStr | Respiratory explants as a model to investigate early events of contagious bovine pleuropneumonia infection |
title_full_unstemmed | Respiratory explants as a model to investigate early events of contagious bovine pleuropneumonia infection |
title_short | Respiratory explants as a model to investigate early events of contagious bovine pleuropneumonia infection |
title_sort | respiratory explants as a model to investigate early events of contagious bovine pleuropneumonia infection |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5766988/ https://www.ncbi.nlm.nih.gov/pubmed/29329577 http://dx.doi.org/10.1186/s13567-017-0500-z |
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