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dc.contributor.authorCheng, Tina
dc.contributor.authorKam, Julia
dc.contributor.authorJohansen, Matt
dc.contributor.authorOehlers, Stefan
dc.date.accessioned2020-02-11T05:58:59Z
dc.date.available2020-02-11T05:58:59Z
dc.date.issued2020-02
dc.identifier.citation129:102782en_AU
dc.identifier.urihttps://hdl.handle.net/2123/21832
dc.description.abstractInfection of zebrafish with natural pathogen Mycobacterium marinum is a useful surrogate for studying the human granulomatous inflammatory response to infection by Mycobacterium tuberculosis. The adaptive immune system of the adult stage zebrafish offers an advance on the commonly used embryo infection model as adult zebrafish form granulomas with striking similarities to human-M. tuberculosis granulomas. Here, we present workflows to perform high content analyses of granulomas in adult zebrafish infected with M. marinum by cryosectioning to take advantage of strong endogenous transgenic fluorescence adapted from common zebrafish embryo infection tools. Specific guides to classifying granuloma necrosis and organisation, quantifying bacterial burden and leukocyte infiltration of granulomas, visualizing foam cell formation, analysing extracellular matrix remodelling and granuloma fibrosis are also provided. We use these methods to characterize neutrophil recruitment to M. marinum granulomas across time and find an inverse relation to granuloma necrosis suggesting granuloma necrosis is not a marker of immunopathology in the natural infection system of the adult zebrafish-M. marinum pairing. The methods can be easily translated to studying the zebrafish adaptive immune response to other chronic and granuloma-forming pathogens.en_AU
dc.description.sponsorshipThis work was supported by an Australian National Health and Medical Research Council CJ Martin Early Career Fellowship APP1053407 and Project Grant APP1099912; The University of Sydney Fellowship G197581; NSW Ministry of Health under the NSW Health Early-Mid Career Fellowships Scheme H18/31086; and the Kenyon Family Foundation Inflammation Award (S.H.O.).en_AU
dc.language.isoenen_AU
dc.publisherMicronen_AU
dc.relationNHMRC APP1053407 and APP1099912en_AU
dc.subjectzebrafishen_AU
dc.subjectmycobacterial infectionen_AU
dc.subjectimmunityen_AU
dc.subjectcryosectionen_AU
dc.subjectfluorescence microscopyen_AU
dc.subjectImageJen_AU
dc.titleHigh content analysis of granuloma histology and neutrophilic inflammation in adult zebrafish infected with Mycobacterium marinumen_AU
dc.typeArticleen_AU
dc.subject.asrcFoR::110802 - Medical Infection Agents (incl. Prions)en_AU
dc.subject.asrcFoR::110707 - Innate Immunityen_AU
dc.identifier.doi10.1016/j.micron.2019.102782
dc.type.pubtypePost-printen_AU


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