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.
Highlights
A type 2 immune response is established in response to stimuli such as allergens and helminth parasites
We have optimised a method for isolating viable lamina propria (LP) leukocytes from the small intestine LP during H.polygyrus infection and we show, using cytokine staining and flow cytometry, that this protocol enables the functional identification of subsets of both the myeloid and CD4+ T cell compartments
Our data demonstrate that we have optimised a protocol that allows for successful isolation and functional characterisation of LP leukocytes from the small intestine of H. polygyrus infected mice at two key timepoints, day 7 and day 14 post-infection
Summary
A type 2 immune response is established in response to stimuli such as allergens and helminth parasites. Murine helminth infections are widely used as models of type 2 immune activity, regulation and disease (Gause et al, 2013). A commonly used helminth is the nematode Heligomosmoides polygyrus, which naturally infects the small intestine of wild mice. This parasite is in the same family of pathogens as the human hookworms Necator americanus and Ancylostoma duodenale, its life cycle is more similar to ruminant parasites such as Haemonchus contortus (Reynolds et al, 2012). The migration of larvae and adult worms through the small intestine wall results in epithelial cell damage. Macrophages and dendritic cells contribute to inflammation and stimulate the activation of Th2 cells, a subset of CD4+ effector T cells
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