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Isolation and functional characterisation of lamina propria leukocytes from helminth-infected, murine small intestine

Holly C. Webster, Anna T. Andrusaite, Amy L. Shergold, Simon W.F. Milling, Georgia Perona-Wright
doi: https://doi.org/10.1101/750786
Holly C. Webster
Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, UK
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Anna T. Andrusaite
Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, UK
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Amy L. Shergold
Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, UK
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Simon W.F. Milling
Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, UK
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Georgia Perona-Wright
Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, UK
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  • For correspondence: georgia.perona-wright@glasgow.ac.uk
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Abstract

The use of helminth infections as tools to understand the type 2 immune response is a well-established technique and important to many areas of immunological research. The phenotype and function of immune cell populations at the site of infection is a key determinant of pathogen clearance. However, infections with helminths such as the murine nematode Heligomosmoides polygryrus cause increased mucus production and thickening of the intestinal wall, which can result in extensive cell death when isolating and analysing cells from the lamina propria (LP). Populations of larger immune cells such as macrophages and dendritic cells are often trapped within mucus or dying tissues. Here we describe an optimised protocol for isolating LP leukocytes from the small intestine of H.polygyrus -infected mice, and we demonstrate phenotypic and functional identification of myeloid and CD4+ T cell subsets using cytokine staining and flow cytometry. Our protocol may provide a useful experimental method for the immunological analysis of the affected tissue site during helminth infections.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted August 29, 2019.
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Isolation and functional characterisation of lamina propria leukocytes from helminth-infected, murine small intestine
Holly C. Webster, Anna T. Andrusaite, Amy L. Shergold, Simon W.F. Milling, Georgia Perona-Wright
bioRxiv 750786; doi: https://doi.org/10.1101/750786
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Isolation and functional characterisation of lamina propria leukocytes from helminth-infected, murine small intestine
Holly C. Webster, Anna T. Andrusaite, Amy L. Shergold, Simon W.F. Milling, Georgia Perona-Wright
bioRxiv 750786; doi: https://doi.org/10.1101/750786

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