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

2020 ◽  
Vol 477 ◽  
pp. 112702 ◽  
Author(s):  
Holly C. Webster ◽  
Anna T. Andrusaite ◽  
Amy L. Shergold ◽  
Simon W.F. Milling ◽  
Georgia Perona-Wright
2019 ◽  
Author(s):  
Holly C. Webster ◽  
Anna T. Andrusaite ◽  
Amy L. Shergold ◽  
Simon W.F. Milling ◽  
Georgia Perona-Wright

AbstractThe 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.


2003 ◽  
Vol 313 (1) ◽  
pp. 47-53 ◽  
Author(s):  
Akira Tamura ◽  
Hiroyuki Soga ◽  
Kei Yaguchi ◽  
Mai Yamagishi ◽  
Takayoshi Toyota ◽  
...  

Author(s):  
D.S. Friend ◽  
N. Ghildyal ◽  
M.F. Gurish ◽  
K.F. Austen ◽  
R.L. Stevens

Trichinella spiralis induces a profound mastocytosis and eosinophilia in the small intestine of the infected mouse. Mouse mast cells (MC) store in their granules various combinations of at least five chymotryptic chymases [designated mouse MC protease (mMCP) 1 to 5], two tryptic proteases designated mMCP-6 and mMCP-7 and an exopeptidase, carboxypeptidase A (mMC-CPA). Using antipeptide, protease -specific antibodies to these MC granule proteases, immunohistochemistry was done to determine the distribution, number and protease phenotype of the MCs in the small intestine and spleen 10 to >60 days after Trichinella infection of BALB/c and C3H mice. TEM was performed to evaluate the granule morphology of the MCs between intestinal epithelial cells and in the lamina propria (mucosal MCs) and in the submucosa, muscle and serosa of the intestine (submucosal MCs).As noted in the table below, the number of submucosal MCs remained constant throughout the study. In contrast, on day 14, the number of MCs in the mucosa increased ~25 fold. Increased numbers of MCs were observed between epithelial cells in the mucosal crypts, in the lamina propria and to a lesser extent, between epithelial cells of the intestinal villi.


2001 ◽  
Vol 120 (5) ◽  
pp. A660-A660
Author(s):  
D MCMICHAEL ◽  
A DAVIES ◽  
E MARSHMAN ◽  
P OTTEWELL ◽  
J JENKINS ◽  
...  

Life Sciences ◽  
2017 ◽  
Vol 169 ◽  
pp. 6-10 ◽  
Author(s):  
Eleanor D. Muise ◽  
Neeru Gandotra ◽  
John J. Tackett ◽  
Michaela C. Bamdad ◽  
Robert A. Cowles

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