High-dimensional analysis of intestinal immune cells during helminth infection

• Published in: eLife 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
• Language: English
• 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
• ISSN: 2050-084X; 2050-084X
• DOI: 10.7554/eLife.51678

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.