RT Journal Article
SR Electronic
T1 Dual RNASeq unveils NTHi-macrophage transcriptomic changes during intracellular persistence
JF European Respiratory Journal
JO Eur Respir J
FD European Respiratory Society
SP 2323
DO 10.1183/13993003.congress-2020.2323
VO 56
IS suppl 64
A1 Jodie Ackland
A1 Ashley Heinson
A1 David Cleary
A1 Myron Christodoulides
A1 Tom Wilkinson
A1 Karl Staples
YR 2020
UL http://erj.ersjournals.com/content/56/suppl_64/2323.abstract
AB 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.FootnotesCite this article as: European Respiratory Journal 2020; 56: Suppl. 64, 2323.This abstract was presented at the 2020 ERS International Congress, in session “Respiratory viruses in the "pre COVID-19" era”.This is an ERS International Congress abstract. No full-text version is available. Further material to accompany this abstract may be available at www.ers-education.org (ERS member access only).