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Dual RNASeq unveils NTHi-macrophage transcriptomic changes during intracellular persistence

Dual RNASeq unveils NTHi-macrophage transcriptomic changes during intracellular persistence
Dual RNASeq unveils NTHi-macrophage transcriptomic changes during intracellular persistence
Nontypeable Haemophilus influenzae (NTHi) is a respiratory tract pathogen associated with severe, neutrophilic asthma. Although macrophages are responsible for orchestrating the immune response and pathogen clearance in the lung, NTHi is able to persist within macrophages. The mechanism of NTHi intracellular persistence is not understood, therefore the aim of this work was to use dual RNASeq to investigate the host-pathogen interactions that allow this persistence. Monocyte-derived macrophages (MDM) were used to model NTHi-macrophage infection. RNA was isolated after 6h and 24h of infection and sequenced using the Illumina NovaSeq 6000 platform. Differential gene expression analysis found expression of 863 MDM genes (FDR p<0.05) conserved across 6h and 24h. Gene set enrichment analysis (GSEA) identified these 863 genes as a core transcriptomic immune response profile, featuring enrichment of defence response and cytokine-mediated signalling pathways. Furthermore, KEGG pathway analysis revealed enriched pathways involved in the response to an intracellular pathogen. In comparison, GSEA of 108 differentially expressed NTHi genes (FDR p<0.05) showed enrichment of ribosome and metabolic pathways, suggesting transcriptomic adaptation of NTHi to intracellular residence within MDM. Taken together, host and pathogen transcriptomic data indicate NTHi intracellular persistence in this model, despite upregulation of macrophage immune response pathways. Ongoing work using lung macrophages from asthma patients will assess whether these gene pathways are detectable and correlate with NTHi persistence in asthma.
0903-1936
Ackland, Jodie
dba59510-7535-47f8-b2ba-2d49dfa7fbd8
Heinson, Ashley
822775d1-9379-4bde-99c3-3c031c3100fb
Cleary, David
f4079c6d-d54b-4108-b346-b0069035bec0
Christodoulides, Myron
eba99148-620c-452a-a334-c1a52ba94078
Wilkinson, Tom
81bda040-30e5-4f86-b7db-db925b936e68
Staples, Karl
e0e9d80f-0aed-435f-bd75-0c8818491fee
Ackland, Jodie
dba59510-7535-47f8-b2ba-2d49dfa7fbd8
Heinson, Ashley
822775d1-9379-4bde-99c3-3c031c3100fb
Cleary, David
f4079c6d-d54b-4108-b346-b0069035bec0
Christodoulides, Myron
eba99148-620c-452a-a334-c1a52ba94078
Wilkinson, Tom
81bda040-30e5-4f86-b7db-db925b936e68
Staples, Karl
e0e9d80f-0aed-435f-bd75-0c8818491fee

Ackland, Jodie, Heinson, Ashley, Cleary, David, Christodoulides, Myron, Wilkinson, Tom and Staples, Karl (2020) Dual RNASeq unveils NTHi-macrophage transcriptomic changes during intracellular persistence. European Respiratory Journal, 56 (Suppl. 64). (doi:10.1183/13993003.congress-2020.2323).

Record type: Meeting abstract

Abstract

Nontypeable Haemophilus influenzae (NTHi) is a respiratory tract pathogen associated with severe, neutrophilic asthma. Although macrophages are responsible for orchestrating the immune response and pathogen clearance in the lung, NTHi is able to persist within macrophages. The mechanism of NTHi intracellular persistence is not understood, therefore the aim of this work was to use dual RNASeq to investigate the host-pathogen interactions that allow this persistence. Monocyte-derived macrophages (MDM) were used to model NTHi-macrophage infection. RNA was isolated after 6h and 24h of infection and sequenced using the Illumina NovaSeq 6000 platform. Differential gene expression analysis found expression of 863 MDM genes (FDR p<0.05) conserved across 6h and 24h. Gene set enrichment analysis (GSEA) identified these 863 genes as a core transcriptomic immune response profile, featuring enrichment of defence response and cytokine-mediated signalling pathways. Furthermore, KEGG pathway analysis revealed enriched pathways involved in the response to an intracellular pathogen. In comparison, GSEA of 108 differentially expressed NTHi genes (FDR p<0.05) showed enrichment of ribosome and metabolic pathways, suggesting transcriptomic adaptation of NTHi to intracellular residence within MDM. Taken together, host and pathogen transcriptomic data indicate NTHi intracellular persistence in this model, despite upregulation of macrophage immune response pathways. Ongoing work using lung macrophages from asthma patients will assess whether these gene pathways are detectable and correlate with NTHi persistence in asthma.

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e-pub ahead of print date: 28 October 2020

Identifiers

Local EPrints ID: 445613
URI: http://eprints.soton.ac.uk/id/eprint/445613
ISSN: 0903-1936
PURE UUID: 7e4ace28-8e7c-445a-958a-2b5e3dd8905f
ORCID for Jodie Ackland: ORCID iD orcid.org/0000-0003-3120-3620
ORCID for David Cleary: ORCID iD orcid.org/0000-0003-4533-0700
ORCID for Myron Christodoulides: ORCID iD orcid.org/0000-0002-9663-4731
ORCID for Karl Staples: ORCID iD orcid.org/0000-0003-3844-6457

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Date deposited: 18 Dec 2020 17:30
Last modified: 07 Oct 2021 02:03

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