High-dimensional analysis of intestinal immune cells during helminth infection

Single cell isolation from helminth-infected murine intestines has been notoriously difficult, due to the strong anti-parasite type 2 immune responses that drive mucus production, tissue remodeling and immune cell infiltration. Through the systematic optimization of a standard intestinal digestion p...

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Bibliographic Details
Published ineLife Vol. 9
Main Authors Ferrer-Font, Laura, Mehta, Palak, Harmos, Phoebe, Schmidt, Alfonso J, Chappell, Sally, Price, Kylie M, Hermans, Ian F, Ronchese, Franca, le Gros, Graham, Mayer, Johannes U
Format Journal Article
LanguageEnglish
Published England eLife Science Publications, Ltd 11.02.2020
eLife Sciences Publications Ltd
eLife Sciences Publications, Ltd
Subjects
Animals
B cells
cell isolation
Confocal microscopy
Dimensional analysis
Duodenum
Duodenum - immunology
Duodenum - parasitology
EDTA
Experiments
Flow cytometry
Flow Cytometry - methods
H. polygyrus
Health aspects
helminth infection
Host-Parasite Interactions - immunology
Immune response
Immunology and Inflammation
Infections
intestinal immune cells
Intestine
Intestines
Isolation
Mice, Inbred C57BL
Microscopy
Nematospiroides dubius
Optimization
Parasites
Protocol
Tools and Resources
type 2 immunity
mouse
helminth infection
inflammation
cell isolation
H. polygyrus
intestinal immune cells
immunology
type 2 immunity
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Summary:Single cell isolation from helminth-infected murine intestines has been notoriously difficult, due to the strong anti-parasite type 2 immune responses that drive mucus production, tissue remodeling and immune cell infiltration. Through the systematic optimization of a standard intestinal digestion protocol, we were able to successfully isolate millions of immune cells from the heavily infected duodenum. To validate that these cells gave an accurate representation of intestinal immune responses, we analyzed them using a high-dimensional spectral flow cytometry panel and confirmed our findings by confocal microscopy. Our cell isolation protocol and high-dimensional analysis allowed us to identify many known hallmarks of anti-parasite immune responses throughout the entire course of helminth infection and has the potential to accelerate single-cell discoveries of local helminth immune responses that have previously been unfeasible.
Bibliography:ObjectType-Article-2
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ISSN:2050-084X
2050-084X
DOI:10.7554/eLife.51678