A Porcine <italic toggle="yes">Ex Vivo</italic> Lung Perfusion Model To Investigate Bacterial Pathogenesis

ABSTRACT The use of animal infection models is essential to understand microbial pathogenesis and to develop and test treatments. Insects and two-dimensional (2D) and 3D tissue models are increasingly being used as surrogates for mammalian models. However, there are concerns about whether these mode...

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Autores principales:	Amy Dumigan, Marianne Fitzgerald, Joana Sá-Pessoa Graca Santos, Umar Hamid, Cecilia M. O’Kane, Danny F. McAuley, Jose A. Bengoechea
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Klebsiella lung infection macrophages pig Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ac8d8fa0631545889e6c7e7bdb4d7659
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spelling	oai:doaj.org-article:ac8d8fa0631545889e6c7e7bdb4d76592021-11-15T15:54:45ZA Porcine <italic toggle="yes">Ex Vivo</italic> Lung Perfusion Model To Investigate Bacterial Pathogenesis10.1128/mBio.02802-192150-7511https://doaj.org/article/ac8d8fa0631545889e6c7e7bdb4d76592019-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02802-19https://doaj.org/toc/2150-7511ABSTRACT The use of animal infection models is essential to understand microbial pathogenesis and to develop and test treatments. Insects and two-dimensional (2D) and 3D tissue models are increasingly being used as surrogates for mammalian models. However, there are concerns about whether these models recapitulate the complexity of host-pathogen interactions. In this study, we developed the ex vivo lung perfusion (EVLP) model of infection using porcine lungs to investigate Klebsiella pneumoniae-triggered pneumonia as a model of respiratory infections. The porcine EVLP model recapitulates features of K. pneumoniae-induced pneumonia lung injury. This model is also useful to assess the pathogenic potential of K. pneumoniae, as we observed that the attenuated Klebsiella capsule mutant strain caused less pathological tissue damage with a concomitant decrease in the bacterial burden compared to that in lungs infected with the wild type. The porcine EVLP model allows assessment of inflammatory responses following infection; similar to the case with the mouse pneumonia model, we observed an increase of il-10 in the lungs infected with the wild type and an increase of ifn-γ in lungs infected with the capsule mutant. This model also allows monitoring of phenotypes at the single-cell level. Wild-type K. pneumoniae skews macrophages toward an M2-like state. In vitro experiments probing pig bone marrow-derived macrophages uncovered the role for the M2 transcriptional factor STAT6 and that Klebsiella-induced il-10 expression is controlled by p38 and extracellular signal-regulated kinase (ERK). Klebsiella-induced macrophage polarization is dependent on the capsule. Together, the findings of this study support the utility of the EVLP model using pig lungs as a platform to investigate the infection biology of respiratory pathogens. IMPORTANCE The implementation of infection models that approximate human disease is essential to understand infections and for testing new therapies before they enter into clinical stages. Rodents are used in most preclinical studies, although the differences between mice and humans have fueled the conclusion that murine studies are unreliable predictors of human outcomes. In this study, we have developed a whole-lung porcine model of infection using the ex vivo lung perfusion (EVLP) system established to recondition human lungs for transplant. As a proof of principle, we provide evidence demonstrating that infection of the porcine EVLP with the human pathogen Klebsiella pneumoniae recapitulates the known features of Klebsiella-triggered pneumonia. Moreover, our data revealed that the porcine EVLP model is useful to reveal features of the virulence of K. pneumoniae, including the manipulation of immune cells. Together, the findings of this study support the utility of the EVLP model using pig lungs as a surrogate host for assessing respiratory infections.Amy DumiganMarianne FitzgeraldJoana Sá-Pessoa Graca SantosUmar HamidCecilia M. O’KaneDanny F. McAuleyJose A. BengoecheaAmerican Society for MicrobiologyarticleKlebsiellalung infectionmacrophagespigMicrobiologyQR1-502ENmBio, Vol 10, Iss 6 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Klebsiella lung infection macrophages pig Microbiology QR1-502
spellingShingle	Klebsiella lung infection macrophages pig Microbiology QR1-502 Amy Dumigan Marianne Fitzgerald Joana Sá-Pessoa Graca Santos Umar Hamid Cecilia M. O’Kane Danny F. McAuley Jose A. Bengoechea A Porcine <italic toggle="yes">Ex Vivo</italic> Lung Perfusion Model To Investigate Bacterial Pathogenesis
description	ABSTRACT The use of animal infection models is essential to understand microbial pathogenesis and to develop and test treatments. Insects and two-dimensional (2D) and 3D tissue models are increasingly being used as surrogates for mammalian models. However, there are concerns about whether these models recapitulate the complexity of host-pathogen interactions. In this study, we developed the ex vivo lung perfusion (EVLP) model of infection using porcine lungs to investigate Klebsiella pneumoniae-triggered pneumonia as a model of respiratory infections. The porcine EVLP model recapitulates features of K. pneumoniae-induced pneumonia lung injury. This model is also useful to assess the pathogenic potential of K. pneumoniae, as we observed that the attenuated Klebsiella capsule mutant strain caused less pathological tissue damage with a concomitant decrease in the bacterial burden compared to that in lungs infected with the wild type. The porcine EVLP model allows assessment of inflammatory responses following infection; similar to the case with the mouse pneumonia model, we observed an increase of il-10 in the lungs infected with the wild type and an increase of ifn-γ in lungs infected with the capsule mutant. This model also allows monitoring of phenotypes at the single-cell level. Wild-type K. pneumoniae skews macrophages toward an M2-like state. In vitro experiments probing pig bone marrow-derived macrophages uncovered the role for the M2 transcriptional factor STAT6 and that Klebsiella-induced il-10 expression is controlled by p38 and extracellular signal-regulated kinase (ERK). Klebsiella-induced macrophage polarization is dependent on the capsule. Together, the findings of this study support the utility of the EVLP model using pig lungs as a platform to investigate the infection biology of respiratory pathogens. IMPORTANCE The implementation of infection models that approximate human disease is essential to understand infections and for testing new therapies before they enter into clinical stages. Rodents are used in most preclinical studies, although the differences between mice and humans have fueled the conclusion that murine studies are unreliable predictors of human outcomes. In this study, we have developed a whole-lung porcine model of infection using the ex vivo lung perfusion (EVLP) system established to recondition human lungs for transplant. As a proof of principle, we provide evidence demonstrating that infection of the porcine EVLP with the human pathogen Klebsiella pneumoniae recapitulates the known features of Klebsiella-triggered pneumonia. Moreover, our data revealed that the porcine EVLP model is useful to reveal features of the virulence of K. pneumoniae, including the manipulation of immune cells. Together, the findings of this study support the utility of the EVLP model using pig lungs as a surrogate host for assessing respiratory infections.
format	article
author	Amy Dumigan Marianne Fitzgerald Joana Sá-Pessoa Graca Santos Umar Hamid Cecilia M. O’Kane Danny F. McAuley Jose A. Bengoechea
author_facet	Amy Dumigan Marianne Fitzgerald Joana Sá-Pessoa Graca Santos Umar Hamid Cecilia M. O’Kane Danny F. McAuley Jose A. Bengoechea
author_sort	Amy Dumigan
title	A Porcine <italic toggle="yes">Ex Vivo</italic> Lung Perfusion Model To Investigate Bacterial Pathogenesis
title_short	A Porcine <italic toggle="yes">Ex Vivo</italic> Lung Perfusion Model To Investigate Bacterial Pathogenesis
title_full	A Porcine <italic toggle="yes">Ex Vivo</italic> Lung Perfusion Model To Investigate Bacterial Pathogenesis
title_fullStr	A Porcine <italic toggle="yes">Ex Vivo</italic> Lung Perfusion Model To Investigate Bacterial Pathogenesis
title_full_unstemmed	A Porcine <italic toggle="yes">Ex Vivo</italic> Lung Perfusion Model To Investigate Bacterial Pathogenesis
title_sort	porcine <italic toggle="yes">ex vivo</italic> lung perfusion model to investigate bacterial pathogenesis
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/ac8d8fa0631545889e6c7e7bdb4d7659
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A Porcine <italic toggle="yes">Ex Vivo</italic> Lung Perfusion Model To Investigate Bacterial Pathogenesis

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