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

Webster, Holly; Andrusaite, Anna; Shergold, Amy L.; Milling, Simon; Perona-Wright, Georgia

doi:10.1016/j.jim.2019.112702

Cited by 13 publications

(17 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For the control group, WT C. reinhardtii algal proteins or PBS/ 2% (v/v) glycerol were added to drinking water at the same volume and timing as the algal TGM1. Mice were sacrificed and isolation of lymphocytes from the Peyer’s patches ( Mosconi et al, 2015 ), mesenteric lymph nodes (MLNs) ( White et al, 2021 ) lamina propria of small ( Webster et al, 2020 ) and large ( Clay et al, 2020 ) intestines were analyzed by flow cytometry.…”

Section: Methodsmentioning

confidence: 99%

Oral delivery of a functional algal-expressed TGF-β mimic halts colitis in a murine DSS model

Smyth

Ren

White

et al. 2021

Journal of Biotechnology

View full text Add to dashboard Cite

Inflammatory bowel disease (IBD) is a set of immunological disorders which can generate chronic pain and fatigue associated with the inflammatory symptoms. The treatment of IBD remains a significant hurdle with current therapies being only partially effective or having significant side effects, suggesting that new therapies that elicit different modes of action and delivery strategies are required. TGM1 is a TGF-β mimic that was discovered from the intestinal helminth parasite Heligmosomoides polygyrus and is thought to be produced by the parasite to suppress the intestinal inflammation response to help evade host immunity, making it an ideal candidate to be developed as a novel anti-inflammatory bio-therapeutic. Here we utilized the expression system of the edible green algae Chlamydomonas reinhardtii in order to recombinantly produce active TGM1 in a form that could be ingested. C. reinhardtii robustly expressed TGM1, and the resultant recombinant protein is biologically active as measured by regulatory T cell induction. When delivered orally to mice, the algal expressed TGM1 is able to ameliorate weight loss, lymphadenopathy, and disease symptoms in a mouse model of DSS-induced colitis, demonstrating the potential of this biologic as a novel treatment of IBD.

show abstract

Section: Methodsmentioning

confidence: 99%

Oral delivery of a functional algal-expressed TGF-β mimic halts colitis in a murine DSS model

Smyth

Ren

White

et al. 2021

Journal of Biotechnology

View full text Add to dashboard Cite

show abstract

“…Published protocols were adapted for isolation of leukocytes from the small-intestine (Webster et al, 2020) and colon (including caecum) (Platt et al, 2010). Both tissues were placed on PBS-soaked paper towel, opened longitudinally (after removal of Peyer’s patches from small intestine), and intestinal contents were scraped off using a metal spatula.…”

Section: Methodsmentioning

confidence: 99%

Spectral flow cytometry reveals metabolic heterogeneity in tissue macrophages

Heieis

Patente

Tak

et al. 2022

Preprint

View full text Add to dashboard Cite

Tissue-macrophage populations are constituted by a mosaic of phenotypes, yet new methods are needed to link metabolic status to the range of phenotypes in vivo. We therefore designed a high-dimensional panel for spectral flow cytometry to investigate the heterogeneity of tissue macrophage metabolism at steady-state, and their metabolic adaptation in response to infection. Distinct metabolic profiles were observed between tissue macrophages from different peripheral organs, as well as within populations from a specific site. As such, our data show multiple metabolic states in macrophages corresponding to relative stages of maturity in both the peritoneal cavity and small intestine. Immune perturbation with helminth infection further showed that peritoneal macrophages acquire an overall highly metabolically active profile, whereas responding intestinal macrophages displayed minimal changes in their metabolic phenotype. Thus, we demonstrate that high-dimensional, flow-based analysis is an exciting method to interrogate the metabolic heterogeneity and dynamics of tissue-macrophage populations.

show abstract

“…Lamina propria leukocytes were isolated as described previously 40 . The MLN was harvested and crushed through a 70μm filter to obtain a single cell suspension.…”

Section: Methodsmentioning

confidence: 99%

Tissue-based IL-10 signalling in helminth infection limits IFNγ expression and promotes the intestinal Th2 response

Webster

Gamino

Shergold

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

Type 2 immunity is activated in response to both allergens and helminth infection. It can be detrimental or beneficial, and there is a pressing need to better understand its regulation. The immunosuppressive cytokine IL-10 is known as a T helper 2 (Th2) effector molecule, but it is currently unclear whether IL-10 dampens or promotes Th2 differentiation during infection. Here we show that helminth infection in mice elicits IL-10 expression in both the intestinal lamina propria and the draining mesenteric lymph node, with higher expression in the infected tissue. In vitro, exogenous IL-10 enhanced Th2 differentiation in isolated CD4+ T cells, increasing expression of GATA3 and production of IL-5 and IL-13. The ability of IL-10 to amplify the Th2 response coincided with its suppression of IFNγ expression and, in vivo, we found that, in intestinal helminth infection, IL-10 receptor expression was higher on Th1 cells in the small intestine than on Th2 cells in the same tissue, or on any Th cell in the draining lymph node. In vivo blockade of IL-10 signalling during helminth infection resulted in an expansion of IFNγ+ and Tbet+ Th1 cells in the small intestine and caused a coincident decrease in IL-13, IL-5 and GATA3 expression by intestinal T cells. Together our data indicate that IL-10 signalling promotes Th2 differentiation during helminth infection at least in part by regulating competing Th1 cells in the infected tissue.

show abstract

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

Cited by 13 publications

References 35 publications

Oral delivery of a functional algal-expressed TGF-β mimic halts colitis in a murine DSS model

Oral delivery of a functional algal-expressed TGF-β mimic halts colitis in a murine DSS model

Spectral flow cytometry reveals metabolic heterogeneity in tissue macrophages

Tissue-based IL-10 signalling in helminth infection limits IFNγ expression and promotes the intestinal Th2 response

Contact Info

Product

Resources

About