US-A transatlantic partnership to identify novel factors that influence the development and function of M cells in the gut epithelium

Lead Research Organisation: University of Edinburgh
Department Name: The Roslin Institute

Abstract

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Description This collaboration grant has enabled us to make some important discoveries on the immunobiology of M cells in the gut epithelium.

Many natural prion diseases are orally acquired. Following exposure via this route, the early replication of some prion strains upon follicular dendritic cells (FDC) in the Peyer's patches in the small intestine is important for the efficient spread of disease to the brain (termed neuroinvasion). However, for the prions to replicate upon the FDC within the Peyer's patches they must first cross the gut lumen. M cells are a unique population of cells within the epithelia overlying the Peyer's patches which are specialised for the transcytosis of particles and microorganisms. Our previous data suggested M cells play an important role in the initial uptake and transfer of prions from the gut lumen into Peyer's patches. Since certain pathogens and inflammatory conditions can dramatically modify the status of M cells in the gut epithelium, is it plausible that such changes may also influence oral prion disease susceptibility. In Peyer's patches stimulation by the cytokine RANKL induces the differentiation of enterocytes into M cells and maintains them in their differentiated state. Therefore, in the current study we manipulated RANKL-stimulation in the gut epithelium to modify M-cell density and studied the effects these changes had on susceptibility to oral prion infection. Mice in which RANK (the receptor for RANKL) is deleted only in the gut epithelium are deficient in M cells and unable to sample particulate antigens from the lumen. We show that in the specific absence of M cells in these mice the uptake of prions from the gut lumen and their replication upon FDC in Peyer's patches are likewise blocked. Conversely, exogenous administration of recombinant RANKL enhances M-cell differentiation within the gut epithelium and significantly enhances the uptake of particulate antigens into Peyer's patches. We also show that recombinant RANKL-mediated induction of M cells enhanced the early uptake of prions into lymphoid tissues and significantly reduced disease susceptibility. Together these data demonstrate that the density of M cells in the epithelia overlying the Peyer's patches in the small intestine has a direct influence on the uptake of prions from the gut lumen and disease susceptibility: in the specific absence of M cells prion disease susceptibility is blocked, whereas their increased density increases disease susceptibility. These data have important implications for our understanding of how changes to the gut epithelium may influence the risk of prion infection.

The transcytosis of antigens across the follicle-associated epithelium (FAE) of Peyer's patches by microfold cells (M cells) is important for the induction of efficient immune responses to mucosal antigens. The mucosal immune response is compromised by ageing, but effects on M cells were unknown. We have shown that M-cell density in the FAE of aged mice was dramatically reduced. As a consequence, aged Peyer's patches were significantly deficient in their ability to transcytose particulate lumenal antigen across the FAE. Ageing specifically impaired the expression of Spi-B and the down-stream functional maturation of M cells. Ageing also dramatically impaired CCL20 expression by the FAE. As a consequence, fewer M-cell-inducing CD11c+ B cells were attracted towards the FAE reducing their ability to promote M-cell maturation. Our data demonstrate that ageing dramatically impedes the functional maturation of M cells, revealing an important ageing-related defect in the mucosal immune system's ability to sample lumenal antigens.
Exploitation Route These data identify an important factor which may influence susceptibility to oral prion infection. We have also identified an important aging related defect in the mucosal immune system. These data will be of significant interest to those interested in the development of mucosal vaccines for use in the elderly.
Sectors Agriculture, Food and Drink,Healthcare
 
Description Determining the role of CSF1R-dependent macrophages in of Paneth cells and the
Amount £520,952 (GBP)
Funding ID MR/S000763/1 
Organisation Medical Research Council (MRC) 
Sector Public
Country United Kingdom
Start 11/2018 
End 10/2021
 
Description Japan Partnering Award: Defining the factors that regulate M cell-development in the intestines of livestock
Amount £42,125 (GBP)
Funding ID BB/S019294/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 06/2019 
End 06/2023
 
Description Study of effects of ageing on M cells
Amount £450,000 (GBP)
Funding ID BB/M024288/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 11/2015 
End 10/2018
 
Title Bacterial 16S rRNA gene metabarcoding of faecal samples from young mice, aged mice and aged mice given young bedding 
Description Project: PRJEB36358 The effects of ageing on M cells and the faecal and caecal microbiota composition in mice The intestinal microbiota helps to maintain the maturation and functioning of the mucosal immune system with its dysbiosis causing detrimental effects. Ageing is known to have a significant impact on the composition of the gut microbiota. Although relatively stable for much of adulthood, ageing induces significant shifts in the intestinal microbiota. Therefore, restoring a "healthy" gut microbiota may have beneficial effects on mucosal immunity. In this study, we studied further the effects of ageing on M-cell status and gut microbiota composition. Faecal samples were collected from aged mice before and at 4 and 6 wk after passive microbiota transfer and from young donor mice. DNA was extracted and prepared for 16S rRNA gene sequencing, targeting the V3 hypervariable region. The sequence files generated with the primers removed are publicly available through the European Nucleotide Archive (ENA) under the project accession number PRJEB36358 https://www.ebi.ac.uk/ena/browser/view/PRJEB36358 
Type Of Material Database/Collection of data 
Year Produced 2020 
Provided To Others? Yes  
Impact These data were included in the following manuscripts: Submitted to Cell Reports (Feb, 2020) Donaldson DS, Pollock J, Vohra P, Stevens MP & Mabbott NA. (2020) Microbial stimulation reverses the age-related decline in M cells in aged mice. Submitted to Cell Reports 17/2/20. Submitted to bioRxiv (Feb, 2020) Donaldson DS, Pollock J, Vohra P, Stevens MP & Mabbott NA. (2020) Microbial stimulation reverses the age-related decline in M cells in aged mice. bioRxiv doi: https://doi.org/10.1101/2020.02.17.943514 
URL https://www.ebi.ac.uk/ena/data/view/PRJEB36358
 
Title Identification of Novel Genes Selectively Expressed in the Follicle-Associated Epithelium from the Meta-Analysis of Transcriptomics Data from Multiple Mouse Cell and Tissue Populations 
Description This study provides new insight into the transcriptome of M cells in the gut epithelium 
Type Of Material Database/Collection of data 
Year Produced 2012 
Provided To Others? Yes  
Impact This study provides new insight into the FAE transcriptome. Further characterization of the candidate genes identified here will aid the identification of novel regulators of cell function in the FAE. 
URL http://www.roslin.ed.ac.uk/neil-mabbott/m-cells-and-the-fae/
 
Description Role of M cells in the sampling of mucosal antigens by B cells in Peyer's patches 
Organisation University of Gothenburg
Department Microbiology and Immunology UGOT
Country Sweden 
Sector Academic/University 
PI Contribution We provided our expertise in M cell immunobiology and use of unique in vivo systems to study M cells
Collaborator Contribution Prof. Nils Lycke is an expert in mucosal immunology and provided his unique systems in which to study the generation of antigen-specific IgA responses in the intestine.
Impact A manuscript was published in Nature Communications (2019) describing how activated Peyer´s patch B cells sample antigen directly from M cells in the subepithelial dome to maintain gut germinal center responses Komban RJ, Stromberg A, Biram A, Cervin J, Lebrero-Fernandez C, Mabbott N, Yrlid U, Shulman Z, Bemark M & Lycke N. (2019) Activated Peyer's patch B cells sample antigen from M cells in the subepithelial dome to maintain gut germinal centre responses. Nature Communications 10:2423.
Start Year 2018
 
Description Study of M cell immunobiology and their role in mucosally-acquired infections (Emory) 
Organisation Emory University
Country United States 
Sector Academic/University 
PI Contribution We provided the ageing and prion disease pathogenesis mouse models
Collaborator Contribution Prof. Ifor Williams provided expertise in M cell immunobiology and mucosal immunology. Prof. Williams also provided unique M-cell-deficient mice (Vil1-Cre Rank-flox mice) as well as reagents to synthesise murine recombinant gst-RANKL. These were invaluable for the study of M cells in mice in vivo.
Impact Several publications: DNA RESEARCH 19, 407-422, (2012); doi:10.1093/dnares/dss022 Mucosal Immunology 5, 216-225; doi: 10.1038/mi.2011.68 Mucosal Immunology 6, 1027-1037; doi:10.1038/mi.2012.141 Mucosal Immunology 6, 666-677; doi:10.1038/mi.2013.30 Nat Nanotechnol. 2015 Apr;10(4):361-9; DOI: 10.1038/NNANO.2015.19 PLoS Pathog 12(12): e1006075; doi:10.1371/journal.ppat.1006075 Prof. WIlliams is also a named collaborator on the following BBSRC grants: BB/J014672/1 BB/M024288/1 BB/K021257/1
Start Year 2009
 
Description Gut cells are gatekeepers of infectious brain diseases, study finds 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact Press release via the media describing our recent research published in PLoS Pathogens
Year(s) Of Engagement Activity 2016
URL http://www.bbsrc.ac.uk/news/health/2016/161214-pr-gut-cells-gatekeepers-of-infectious-brain-diseases...
 
Description Keeping bugs at Bay: a Public Engagement Activity at the Roslin Institute 2014 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact This activity aims to teach people how the immune system fights bugs.
Year(s) Of Engagement Activity 2014
 
Description Keeping bugs at Bay: a Public Engagement Activity at the Royal Highland Show 2015 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Industry/Business
Results and Impact This activity aims to teach people how the immune system fights bugs.
Year(s) Of Engagement Activity 2015
 
Description Scientific Writing Workshop at the Young Microbiologists Symposium, Dundee, 2016 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact The journal PLoS Pathogens invited me to host a scientific writing workshop aimed at junior research scientists such as new post-docs or PhD students
Year(s) Of Engagement Activity 2016
URL http://www.lifesci.dundee.ac.uk/other/yms/