Interaction of Klebsiella pneumoniae with tissue macrophages in a mouse infection model and ex-vivo pig organ perfusions: an exploratory investigation

Summary: Background: Hypervirulent Klebsiella pneumoniae (hvKp) strains of capsule type K1 and K2 cause invasive infections associated with hepatic abscesses, which can be difficult to treat and are frequently associated with relapsing infections. Other K pneumoniae strains (non-hvKp), including li...

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Autores principales:	Joseph J Wanford, PhD, Ryan G Hames, BSc, David Carreno, PhD, Zydrune Jasiunaite, BSc, Wen Y Chung, PhD, Fabio Arena, MD, Vincenzo Di Pilato, PhD, Kornelis Straatman, PhD, Kevin West, ProfPhD, Robeena Farzand, PhD, Mariagrazia Pizza, MA, Luisa Martinez-Pomares, PhD, Peter W Andrew, ProfPhD, E Richard Moxon, ProfFRS, Ashley R Dennison, ProfMD, Gian Maria Rossolini, ProfMD, Marco R Oggioni, ProfMD
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Medicine (General) R5-920 Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/98b688093a574746b4bdada9d71052cd
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spelling	oai:doaj.org-article:98b688093a574746b4bdada9d71052cd2021-12-02T05:03:51ZInteraction of Klebsiella pneumoniae with tissue macrophages in a mouse infection model and ex-vivo pig organ perfusions: an exploratory investigation2666-524710.1016/S2666-5247(21)00195-6https://doaj.org/article/98b688093a574746b4bdada9d71052cd2021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2666524721001956https://doaj.org/toc/2666-5247Summary: Background: Hypervirulent Klebsiella pneumoniae (hvKp) strains of capsule type K1 and K2 cause invasive infections associated with hepatic abscesses, which can be difficult to treat and are frequently associated with relapsing infections. Other K pneumoniae strains (non-hvKp), including lineages that have acquired carbapenem resistance, do not manifest this pathology. In this work we aimed to test the hypothesis that within-macrophage replication is a key mechanism underpinning abscess formation in hvKp infections. Methods: In this exploratory investigation, to study the pathophysiology of abscess formation, mice were intravenously infected with 106 colony forming units (CFU) of either hvKp isolates (six strains) or non-hvKp isolates (seven strains). Intracellular bacterial replication and neutrophil influx in liver and spleen was quantified by fluorescence microscopy of sliced cryopreserved organs of mice collected 30 min, 6 h, and 24 h after infection with the aim to provide data of bacterial association to Kupffer cells in the liver and to the different tissue macrophages in the spleen. Microbiological and microscopy analysis of an ex-vivo model of pig liver and spleen infection were used to confirm within-macrophage replication. Pig organs were perfused with heparinised, autologous pig's blood and injected with 6·5 × 107 CFU of hvKp K2 sequence type 25 strain GMR151. Blood and tissue biopsies collected before infection and 30 min, 1 h, 2 h, 3 h, 4 h, and 5 h after infection were used to measure bacterial counts and to identify the subcellular localisation of bacteria by immunohistochemistry analysis. Findings: We show that hvKp resisted phagocyte-mediated clearance and replicated in mouse liver macrophages to form clusters 6 h after infection, with a mean of 7·0 bacteria per Kupffer cell (SD 6·2); however, non-hvKp were efficiently cleared (mean 1·5 bacteria per cell [SD 1·1]). HvKp infection promoted neutrophil recruitment to sites of infection, which in the liver resulted in histopathological signs of abscess formation as early as 24 h post-infection. Experiments in pig organs which share a high functional and anatomical resemblance to human organs, provided strong evidence for the propensity of hvKp to replicate within the hepatic macrophages. Interpretation: These findings show subversion of innate immune processes in the liver by K pneumoniae and resistance to Kupffer cell mediated clearance as an explanation for the propensity of hvKp strains to cause hepatic abscesses. Funding: University of Oxford and a Royal Society Wolfson grant funded biosafety facility.Joseph J Wanford, PhDRyan G Hames, BScDavid Carreno, PhDZydrune Jasiunaite, BScWen Y Chung, PhDFabio Arena, MDVincenzo Di Pilato, PhDKornelis Straatman, PhDKevin West, ProfPhDRobeena Farzand, PhDMariagrazia Pizza, MALuisa Martinez-Pomares, PhDPeter W Andrew, ProfPhDE Richard Moxon, ProfFRSAshley R Dennison, ProfMDGian Maria Rossolini, ProfMDMarco R Oggioni, ProfMDElsevierarticleMedicine (General)R5-920MicrobiologyQR1-502ENThe Lancet Microbe, Vol 2, Iss 12, Pp e695-e703 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920 Microbiology QR1-502
spellingShingle	Medicine (General) R5-920 Microbiology QR1-502 Joseph J Wanford, PhD Ryan G Hames, BSc David Carreno, PhD Zydrune Jasiunaite, BSc Wen Y Chung, PhD Fabio Arena, MD Vincenzo Di Pilato, PhD Kornelis Straatman, PhD Kevin West, ProfPhD Robeena Farzand, PhD Mariagrazia Pizza, MA Luisa Martinez-Pomares, PhD Peter W Andrew, ProfPhD E Richard Moxon, ProfFRS Ashley R Dennison, ProfMD Gian Maria Rossolini, ProfMD Marco R Oggioni, ProfMD Interaction of Klebsiella pneumoniae with tissue macrophages in a mouse infection model and ex-vivo pig organ perfusions: an exploratory investigation
description	Summary: Background: Hypervirulent Klebsiella pneumoniae (hvKp) strains of capsule type K1 and K2 cause invasive infections associated with hepatic abscesses, which can be difficult to treat and are frequently associated with relapsing infections. Other K pneumoniae strains (non-hvKp), including lineages that have acquired carbapenem resistance, do not manifest this pathology. In this work we aimed to test the hypothesis that within-macrophage replication is a key mechanism underpinning abscess formation in hvKp infections. Methods: In this exploratory investigation, to study the pathophysiology of abscess formation, mice were intravenously infected with 106 colony forming units (CFU) of either hvKp isolates (six strains) or non-hvKp isolates (seven strains). Intracellular bacterial replication and neutrophil influx in liver and spleen was quantified by fluorescence microscopy of sliced cryopreserved organs of mice collected 30 min, 6 h, and 24 h after infection with the aim to provide data of bacterial association to Kupffer cells in the liver and to the different tissue macrophages in the spleen. Microbiological and microscopy analysis of an ex-vivo model of pig liver and spleen infection were used to confirm within-macrophage replication. Pig organs were perfused with heparinised, autologous pig's blood and injected with 6·5 × 107 CFU of hvKp K2 sequence type 25 strain GMR151. Blood and tissue biopsies collected before infection and 30 min, 1 h, 2 h, 3 h, 4 h, and 5 h after infection were used to measure bacterial counts and to identify the subcellular localisation of bacteria by immunohistochemistry analysis. Findings: We show that hvKp resisted phagocyte-mediated clearance and replicated in mouse liver macrophages to form clusters 6 h after infection, with a mean of 7·0 bacteria per Kupffer cell (SD 6·2); however, non-hvKp were efficiently cleared (mean 1·5 bacteria per cell [SD 1·1]). HvKp infection promoted neutrophil recruitment to sites of infection, which in the liver resulted in histopathological signs of abscess formation as early as 24 h post-infection. Experiments in pig organs which share a high functional and anatomical resemblance to human organs, provided strong evidence for the propensity of hvKp to replicate within the hepatic macrophages. Interpretation: These findings show subversion of innate immune processes in the liver by K pneumoniae and resistance to Kupffer cell mediated clearance as an explanation for the propensity of hvKp strains to cause hepatic abscesses. Funding: University of Oxford and a Royal Society Wolfson grant funded biosafety facility.
format	article
author	Joseph J Wanford, PhD Ryan G Hames, BSc David Carreno, PhD Zydrune Jasiunaite, BSc Wen Y Chung, PhD Fabio Arena, MD Vincenzo Di Pilato, PhD Kornelis Straatman, PhD Kevin West, ProfPhD Robeena Farzand, PhD Mariagrazia Pizza, MA Luisa Martinez-Pomares, PhD Peter W Andrew, ProfPhD E Richard Moxon, ProfFRS Ashley R Dennison, ProfMD Gian Maria Rossolini, ProfMD Marco R Oggioni, ProfMD
author_facet	Joseph J Wanford, PhD Ryan G Hames, BSc David Carreno, PhD Zydrune Jasiunaite, BSc Wen Y Chung, PhD Fabio Arena, MD Vincenzo Di Pilato, PhD Kornelis Straatman, PhD Kevin West, ProfPhD Robeena Farzand, PhD Mariagrazia Pizza, MA Luisa Martinez-Pomares, PhD Peter W Andrew, ProfPhD E Richard Moxon, ProfFRS Ashley R Dennison, ProfMD Gian Maria Rossolini, ProfMD Marco R Oggioni, ProfMD
author_sort	Joseph J Wanford, PhD
title	Interaction of Klebsiella pneumoniae with tissue macrophages in a mouse infection model and ex-vivo pig organ perfusions: an exploratory investigation
title_short	Interaction of Klebsiella pneumoniae with tissue macrophages in a mouse infection model and ex-vivo pig organ perfusions: an exploratory investigation
title_full	Interaction of Klebsiella pneumoniae with tissue macrophages in a mouse infection model and ex-vivo pig organ perfusions: an exploratory investigation
title_fullStr	Interaction of Klebsiella pneumoniae with tissue macrophages in a mouse infection model and ex-vivo pig organ perfusions: an exploratory investigation
title_full_unstemmed	Interaction of Klebsiella pneumoniae with tissue macrophages in a mouse infection model and ex-vivo pig organ perfusions: an exploratory investigation
title_sort	interaction of klebsiella pneumoniae with tissue macrophages in a mouse infection model and ex-vivo pig organ perfusions: an exploratory investigation
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/98b688093a574746b4bdada9d71052cd
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Interaction of Klebsiella pneumoniae with tissue macrophages in a mouse infection model and ex-vivo pig organ perfusions: an exploratory investigation

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