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Characterization of macrophage infiltration and polarization by double fluorescence immunostaining in mouse colonic mucosa
Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR). Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine).
Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR). Umeå University, Faculty of Science and Technology, Department of Molecular Biology (Faculty of Science and Technology).
Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR). Umeå University, Faculty of Science and Technology, Department of Molecular Biology (Faculty of Science and Technology).ORCID iD: 0000-0002-1209-0942
Univ. Bordeaux, INSERM, BaRITOn, U1053, Bordeaux, France.
2021 (English)In: STAR Protocols, E-ISSN 2666-1667, Vol. 2, no 4, article id 100833Article in journal (Refereed) Published
Abstract [en]

We recently characterized the association between DNA damage and immunoresponse in vivo in colonic mucosa of mice infected with a Salmonella Typhimurium strain expressing a genotoxin, known as typhoid toxin. In this protocol, we describe how to assess the extent and features of infiltrating macrophages by double immunofluorescence. Total macrophage population was determined using an F4/80 antibody, whereas the specific M2-like population was assessed using a CD206 antibody. For complete details on the use and execution of this protocol, please refer to Martin et al. (2021).

Place, publisher, year, edition, pages
Cell Press , 2021. Vol. 2, no 4, article id 100833
Keywords [en]
Cell-based assays, In situ hybridization, Microscopy
National Category
Immunology
Identifiers
URN: urn:nbn:se:umu:diva-202969DOI: 10.1016/j.xpro.2021.100833Scopus ID: 2-s2.0-85122830767OAI: oai:DiVA.org:umu-202969DiVA, id: diva2:1727007
Funder
Swedish Cancer Society, 20 0699 PjFSwedish Cancer Society, CAN 2017/315Swedish Research Council, 2018–02521Umeå UniversityThe Kempe Foundations, JCK-1826Available from: 2023-01-14 Created: 2023-01-14 Last updated: 2023-03-24Bibliographically approved
In thesis
1. Effects of bacterial genotoxins on immune modulation, chronic inflammation and cancer development
Open this publication in new window or tab >>Effects of bacterial genotoxins on immune modulation, chronic inflammation and cancer development
2023 (English)Doctoral thesis, comprehensive summary (Other academic)
Alternative title[sv]
Effekter av bakteriella genotoxiner på immunmodulering, kronisk inflammation och cancerutveckling
Abstract [en]

The intestinal microbiome of Inflammatory Bowel Disease and colorectal cancer patients is enriched in genotoxin-producing bacteria, which cause DNA damage in the host cells.

Genotoxins have recently been identified as a novel family of effectors produced by pathogenic and commensal bacteria. At present, only three types of bacterial genotoxins have been identified: colibactin, produced by some Escherichia coli strains; cytolethal distending toxins, produced by several Gram-negative pathogens; and the typhoid toxin, produced by Salmonella enterica serovar Typhi.

Exposure to high toxin doses activates the classical DNA damage response, which consequently blocks proliferation and eventually induces death in mammalian cells. However, exposure to low toxin doses has shown to promote classical signs of carcinogenesis in vitro, such as cell survival and acquisition of genomic instability. Despite an extensive characterization of their mode of action in vitro, we have a poor understanding of genotoxins´ role in chronic infection and, considering the genotoxic potential, of their carcinogenic capacity. To investigate further the role played by the genotoxins, we focused specifically on Salmonella Typhi, since it is the only genotoxin-producing bacterium that induces a chronic infection associated with increased risk of tumor development in humans. 

The results presented in this thesis show that these unusual bacterial effectors are not classical toxins, but rather act as immunomodulators, highlighting a complex and tissue-specific crosstalk between two highly conserved stress responses: the immune response and the DNA damage response. 

Our data indicate that the impact of genotoxin-producing bacteria on the modulation of the host mucosal response is still poorly characterized and suggest that the host-microbe interaction and the tissue microenvironment are the key players in determining the outcome of the infection and the toxin carcinogenic potential. 

Place, publisher, year, edition, pages
Umeå: Umeå University, 2023. p. 93
National Category
Immunology Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject
Immunology
Identifiers
urn:nbn:se:umu:diva-203905 (URN)978-91-7855-971-8 (ISBN)978-91-7855-972-5 (ISBN)
Public defence
2023-02-24, Major Groove, Department of Molecular Biology, University hospital area, building 6L., Umeå, 09:00 (English)
Opponent
Supervisors
Available from: 2023-02-03 Created: 2023-01-23 Last updated: 2023-01-25Bibliographically approved

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Lopez Chiloeches, MariaBergonzini, AnnaFrisan, Teresa

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Lopez Chiloeches, MariaBergonzini, AnnaFrisan, Teresa
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Umeå Centre for Microbial Research (UCMR)Department of Molecular Biology (Faculty of Medicine)Department of Molecular Biology (Faculty of Science and Technology)
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