Dual RNA-seq in <named-content content-type="genus-species">Streptococcus pneumoniae</named-content> Infection Reveals Compartmentalized Neutrophil Responses in Lung and Pleural Space

ABSTRACT Streptococcus pneumoniae is the dominant cause of community-acquired pneumonia worldwide. Invasion of the pleural space is common and results in increased mortality. We set out to determine the bacterial and host factors that influence invasion of the pleural space. In a murine model of pne...

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Autores principales:	Neil D. Ritchie, Tom J. Evans
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	transcriptomics dual RNA-seq neutrophils pneumonia Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/3775e810f3594c339db740d5e129bbd0
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spelling	oai:doaj.org-article:3775e810f3594c339db740d5e129bbd02021-12-02T18:39:47ZDual RNA-seq in <named-content content-type="genus-species">Streptococcus pneumoniae</named-content> Infection Reveals Compartmentalized Neutrophil Responses in Lung and Pleural Space10.1128/mSystems.00216-192379-5077https://doaj.org/article/3775e810f3594c339db740d5e129bbd02019-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSystems.00216-19https://doaj.org/toc/2379-5077ABSTRACT Streptococcus pneumoniae is the dominant cause of community-acquired pneumonia worldwide. Invasion of the pleural space is common and results in increased mortality. We set out to determine the bacterial and host factors that influence invasion of the pleural space. In a murine model of pneumococcal infection, we isolated neutrophil-dominated samples of bronchoalveolar and pleural fluid containing bacteria 48 hours after infection. Using dual RNA sequencing (RNA-seq), we characterized bacterial and host transcripts that were differentially regulated between these compartments and bacteria in broth and resting neutrophils, respectively. Pleural and lung samples showed upregulation of genes involved in the positive regulation of neutrophil extravasation but downregulation of genes mediating bacterial killing. Compared to the lung samples, cells within the pleural space showed marked upregulation of many genes induced by type I interferons, which are cytokines implicated in preventing bacterial transmigration across epithelial barriers. Differences in the bacterial transcripts between the infected samples and bacteria grown in broth showed the upregulation of genes in the bacteriocin locus, the pneumococcal surface adhesin PsaA, and the glycopeptide resistance gene vanZ; the gene encoding the ClpP protease was downregulated in infection. One hundred sixty-nine intergenic putative small bacterial RNAs were also identified, of which 43 (25.4%) small RNAs had been previously described. Forty-two of the small RNAs were upregulated in pleura compared to broth, including many previously identified as being important in virulence. Our results have identified key host and bacterial responses to invasion of the pleural space that can be potentially exploited to develop alternative antimicrobial strategies for the prevention and treatment of pneumococcal pleural disease. IMPORTANCE The factors that regulate the passage of bacteria between different anatomical compartments are unclear. We have used an experimental model of infection with Streptococcus pneumoniae to examine the host and bacterial factors involved in the passage of bacteria from the lung to the pleural space. The transcriptional profile of host and bacterial cells within the pleural space and lung was analyzed using deep sequencing of the entire transcriptome using the technique of dual RNA-seq. We found significant differences in the host and bacterial RNA profiles in infection, which shed light on the key factors that allow passage of this bacterium into the pleural space.Neil D. RitchieTom J. EvansAmerican Society for Microbiologyarticletranscriptomicsdual RNA-seqneutrophilspneumoniaMicrobiologyQR1-502ENmSystems, Vol 4, Iss 4 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	transcriptomics dual RNA-seq neutrophils pneumonia Microbiology QR1-502
spellingShingle	transcriptomics dual RNA-seq neutrophils pneumonia Microbiology QR1-502 Neil D. Ritchie Tom J. Evans Dual RNA-seq in <named-content content-type="genus-species">Streptococcus pneumoniae</named-content> Infection Reveals Compartmentalized Neutrophil Responses in Lung and Pleural Space
description	ABSTRACT Streptococcus pneumoniae is the dominant cause of community-acquired pneumonia worldwide. Invasion of the pleural space is common and results in increased mortality. We set out to determine the bacterial and host factors that influence invasion of the pleural space. In a murine model of pneumococcal infection, we isolated neutrophil-dominated samples of bronchoalveolar and pleural fluid containing bacteria 48 hours after infection. Using dual RNA sequencing (RNA-seq), we characterized bacterial and host transcripts that were differentially regulated between these compartments and bacteria in broth and resting neutrophils, respectively. Pleural and lung samples showed upregulation of genes involved in the positive regulation of neutrophil extravasation but downregulation of genes mediating bacterial killing. Compared to the lung samples, cells within the pleural space showed marked upregulation of many genes induced by type I interferons, which are cytokines implicated in preventing bacterial transmigration across epithelial barriers. Differences in the bacterial transcripts between the infected samples and bacteria grown in broth showed the upregulation of genes in the bacteriocin locus, the pneumococcal surface adhesin PsaA, and the glycopeptide resistance gene vanZ; the gene encoding the ClpP protease was downregulated in infection. One hundred sixty-nine intergenic putative small bacterial RNAs were also identified, of which 43 (25.4%) small RNAs had been previously described. Forty-two of the small RNAs were upregulated in pleura compared to broth, including many previously identified as being important in virulence. Our results have identified key host and bacterial responses to invasion of the pleural space that can be potentially exploited to develop alternative antimicrobial strategies for the prevention and treatment of pneumococcal pleural disease. IMPORTANCE The factors that regulate the passage of bacteria between different anatomical compartments are unclear. We have used an experimental model of infection with Streptococcus pneumoniae to examine the host and bacterial factors involved in the passage of bacteria from the lung to the pleural space. The transcriptional profile of host and bacterial cells within the pleural space and lung was analyzed using deep sequencing of the entire transcriptome using the technique of dual RNA-seq. We found significant differences in the host and bacterial RNA profiles in infection, which shed light on the key factors that allow passage of this bacterium into the pleural space.
format	article
author	Neil D. Ritchie Tom J. Evans
author_facet	Neil D. Ritchie Tom J. Evans
author_sort	Neil D. Ritchie
title	Dual RNA-seq in <named-content content-type="genus-species">Streptococcus pneumoniae</named-content> Infection Reveals Compartmentalized Neutrophil Responses in Lung and Pleural Space
title_short	Dual RNA-seq in <named-content content-type="genus-species">Streptococcus pneumoniae</named-content> Infection Reveals Compartmentalized Neutrophil Responses in Lung and Pleural Space
title_full	Dual RNA-seq in <named-content content-type="genus-species">Streptococcus pneumoniae</named-content> Infection Reveals Compartmentalized Neutrophil Responses in Lung and Pleural Space
title_fullStr	Dual RNA-seq in <named-content content-type="genus-species">Streptococcus pneumoniae</named-content> Infection Reveals Compartmentalized Neutrophil Responses in Lung and Pleural Space
title_full_unstemmed	Dual RNA-seq in <named-content content-type="genus-species">Streptococcus pneumoniae</named-content> Infection Reveals Compartmentalized Neutrophil Responses in Lung and Pleural Space
title_sort	dual rna-seq in <named-content content-type="genus-species">streptococcus pneumoniae</named-content> infection reveals compartmentalized neutrophil responses in lung and pleural space
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/3775e810f3594c339db740d5e129bbd0
work_keys_str_mv	AT neildritchie dualrnaseqinnamedcontentcontenttypegenusspeciesstreptococcuspneumoniaenamedcontentinfectionrevealscompartmentalizedneutrophilresponsesinlungandpleuralspace AT tomjevans dualrnaseqinnamedcontentcontenttypegenusspeciesstreptococcuspneumoniaenamedcontentinfectionrevealscompartmentalizedneutrophilresponsesinlungandpleuralspace
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Dual RNA-seq in <named-content content-type="genus-species">Streptococcus pneumoniae</named-content> Infection Reveals Compartmentalized Neutrophil Responses in Lung and Pleural Space

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