| Title: |
Disease-attenuated pneumococcal biosynthesis gene mutants invade the mucosal epithelium and induce innate immunity |
| Authors: |
Weight, Caroline; Pollara, Gabriele; Betts, Modupeh; Ragazzini, Roberta; Ramos-Sevillano, Elisa; Reiné, Jesús; Whelan, Matthew; Guerra-Assunção, José Afonso; Connor, Michael; Bonfanti, Paola; Jolly, Clare; Noursadeghi, Mahdad; Ferreira, Daniela; Brown, Jeremy; Heyderman, Robert |
| Contributors: |
University College London UCL (UCL); The Francis Crick Institute London; Oxford Vaccine Group; University of Oxford-Oxford NIHR Biomedical Research Centre-The Churchill hospital; Liverpool School of Tropical Medicine (LSTM); Chromatine et Infection - Chromatin and Infection; Institut Pasteur Paris (IP)-Université Paris Cité (UPCité); This study was funded by CMW from a Postdoctoral Innovation Award from UCL, a Pump-Priming Award and Enhancement Award from the Human Infection Challenge Network for Vaccine Development (HIC-VAC (funded by the GCRF Networks in Vaccines Research and Development) which is co-funded by the MRC and BBSRC). This UK funded award is part of the EDCTP2 programme supported by the European Union. CMW and RSH are supported by the Medical Research Council (grant MR/T016329/1). The authors would like to thank Prof. Peter Hermans (Radboud University Medical Centre) for donating the 6B pneumococcal strain originally from Prof.Birgitta Henriques-Normark (Karolinska Institute) and obtained directly from DF at LSTM. EHPC sample collection was supported by the Medical Research Council (grant MR/M011569/1), Bill and Melinda Gates Foundation (grant OPP1117728) awarded to DF and the National Institute for Health Research (NIHR) Local Comprehensive Research Network. The authors also wish to thank the EHPC Clinical Team at LSTM and all the volunteers. Confocal imaging facilities at LSTM were funded by a Wellcome Trust Multi-User Equipment Grant (104936/Z/14/Z). Electron microscopy processing and image capture was performed by Dr. Elizabeth Slavik-Smith in the Biosciences EMfacility at UCL. MC is supported by a Springboard to Independence grant (AirwayStasis) from the French Government’s Investissement d’Avenir program, the Laboratoire d’Excellence ‘‘Integrative Biology of Emerging Infectious Diseases” (ANR-10-LABX-62-IBEID) and the Chromatin and infection unit headed by Melanie A. Hamon. RNA-Seq library preparation was undertaken at UCL through the UCL/UCLH Biomedical Research Centre and MRC funded Pathogen Genomics Unit (PGU, G0900950). All RNAseq data processing was completed by the PGU. MN is supported by the Wellcome Trust and by NIHR Biomedical Research Centre Funding to University College HospitalsNHS Foundation Trust and University College London. RR is supported by Marie Skłodowska-Curie Individual Fellowships 896014. PB is supported by the European Research Council (ERC-Stg No.639429), the Rosetrees Trust (M362-F1; M553), the CF Trust (SRC006; SRC020), the London Advanced Therapies - Research England (C2N-AT.006), the MRC Confidence in Concept scheme (Award: MC_PC_17180), the Innovate UK (Smart grant n.10005465), the Duchenne Parent Project (DPP) and the NIHR GOSH BRC. MW is supported by a 223065 Wellcome Investigator Award to Clare Jolly, UCL. ERS and JSB are supported by MRC grant R/N02687X/1 and Wellcome grant 221803/Z/20/Z. JSB, MN, and RSH acknowledge funding from the Department of Health’s National Institute of Health and Care Research Biomedical Research Centre funding to UCL and UCLH. RSHis a NIHR Senior Investigator.; ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010); European Project: 896014,H2020-MSCA-IF-2019,H2020-MSCA-IF-2019,ThymEForT(2021); European Project: 639429,ERC-2014-STG,ERC-2014-STG,Rethyming(2015) |
| Source: |
https://hal.science/hal-05121334 ; 2025. |
| Publisher Information: |
CCSD |
| Publication Year: |
2025 |
| Subject Terms: |
Innate immunity; disease; colonisation; airway epithelium; Streptococcus pneumoniae; host-pathogen interactions; Biosynthesis genes; [SDV]Life Sciences [q-bio] |
| Description: |
International audience ; Nasopharyngeal colonisation by Streptococcus pneumoniae is characterised by adherence to epithelial cells, microinvasion and innate immune activation. Previously, we have shown that two biosynthesis gene mutants of S. pneumoniae serotype 6B strain ( ΔproABC and Δfhs with an additional ΔpiaA mutation) can successfully colonise murine and human models without causing disease. Here, we test the hypothesis that epithelial microinvasion and an innate immune response persist despite disease attenuation. We show that although the ΔproABC and Δfhs mutations do not attenuate microinvasion in either experimental human pneumococcal challenge or infection of epithelial cell models, there was less transmigration of Detroit 562 nasopharyngeal epithelial cells by the biosynthesis gene mutants than the WT. Cellular reorganisation by primary human airway epithelial cells varied considerably between strains. Compared to the WT, infection of Detroit 562 epithelial cells by the Δfhs/piaA mutant but not the ΔproABC/piaA mutant was less pro-inflammatory, induced less caspase 8 production, associated increased pneumococcal hydrogen peroxide secretion and reduced pneumolysin activity. Under serum stress, these biosynthesis gene mutations had a broad impact on the expression of pneumococcal virulence genes (e.g. ply , nanA and psaA ), those regulating oxidative stress (e.g. SpxB , lctO and adhE ), and genes involved in purine and carbohydrate metabolism. However, although these may result in disease attenuation, they were not directly linked to effects on microinvasion, cellular reorganisation or the epithelial innate immune response. These findings suggest that the strain differences observed were driven by the differential expression of multiple bacterial virulence and metabolic pathways, rather than any single gene or pathway of genes. These data highlight the complex impact of single gene mutations on bacterial virulence and suggest that the virulence determinants of pneumococcal epithelial colonisation, ... |
| Document Type: |
report |
| Language: |
English |
| Relation: |
https://doi.org/10.5281/zenodo.7997789; info:eu-repo/grantAgreement//896014/EU/Dissecting the role of thymic epithelial cells in T lymphocytes maturation during human foetal development/ThymEForT; info:eu-repo/grantAgreement//639429/EU/Rebuilding the human thymus to create a tolerising system for allogeneic tissue and organ transplantation/Rethyming; BIORXIV: 2023.06.15.545009 |
| DOI: |
10.1101/2023.06.15.545009 |
| Availability: |
https://hal.science/hal-05121334; https://doi.org/10.1101/2023.06.15.545009 |
| Accession Number: |
edsbas.74BC5E2C |
| Database: |
BASE |