Affiliations:
1Translational Medicine Program, Research Institute,
Hospital for Sick Children, Toronto, Ontario, Canada
2Department of Immunology, Temerty Faculty of
Medicine, University of Toronto, Toronto, Ontario, Canada
3Centre for Computational Medicine, Research
Institute, Hospital for Sick Children, Toronto, Ontario, Canada
4Division of Advanced Diagnostics, Toronto General
Hospital Research Institute, University Health Network, Toronto,
Ontario, Canada
5Institute of Medical Science, Temerty Faculty of
Medicine, University of Toronto, Toronto, Ontario, Canada
6Department of Preclinical Pharmacology and In-Vitro
Toxicology, Fraunhofer Institute for Toxicology and Experimental
Medicine, Hannover, Germany
7Division of Immunology and Allergy, SickKids Food
Allergy and Anaphylaxis Program, Hospital for Sick Children, Toronto,
Ontario, Canada
8Department of Paediatrics, Temerty Faculty of
Medicine, University of Toronto, Toronto, Ontario, Canada
9National Food Institute, Technical University of
Denmark, Kgs. Lyngby, Denmark
10Department of Laboratory Medicine and Pathobiology,
Temerty Faculty of Medicine, University of Toronto, Toronto, Ontario,
Canada
11Department of Paediatric Laboratory Medicine, The
Hospital for Sick Children, Toronto, Ontario, Canada
12Surgical Pathology, University Health Network,
Toronto, Ontario, Canada
13Surgical Oncology and Minimally Invasive Surgery,
Princess Margaret Cancer Centre, University Health Network, Toronto,
Ontario, Canada
14Department of Surgery, Temerty Faculty of Medicine,
University of Toronto, Toronto, Ontario, Canada
15Division of General and Thoracic Surgery, Hospital
for Sick Children, Toronto, Ontario, Canada
16Karl Landsteiner University of Health Sciences,
Krems an der Donau, Austria
17Department of Pediatric and Adolescent Medicine,
University Hospital St. Pölten, St. Pölten, Austria
Corresponding Author : Thomas Eiwegger, MD, Department of
Pediatric and Adolescent Medicine, University Hospital St. Pölten,
Dunant-Platz 1, 3100 St. Pölten, Austria. E-mail:
thomas.eiwegger@stpoelten.lknoe.at, Tel.: +43 2742-9004-11740
Conflict of Interest Statement: LH, AC, XY, KY, AB, AK, HO,
KLB, GRS, IS, MG, FAQ, KS, and PPLC have nothing to disclose. JEMU
reports grants/research support from DBV Technologies, Regeneron, CIHR,
ALK-Abelló, SickKids Food Allergy and Anaphylaxis Program, Advisory
board for Pfizer, Kaleo, Bausch Health, Food Allergy Canada; in-kind
drug donation from Novartis, other for Astra-Zeneca, all outside the
current work. TE reports to act as local PI for company sponsored trials
by DBV Therapeutics, Greer Stallergens, and sub-investigator for
Regeneron and ALK-Abelló. He is Co-Investigator or scientific lead in
three investigator-initiated oral immunotherapy trials supported by the
SickKids Food Allergy and Anaphylaxis Program and serves as an associate
editor for Allergy. He/his lab received unconditional/in-kind
contributions from Macro Array Diagnostics and an unrestricted grant
from ALK-Abelló. He holds advisory board roles for ALK-Abelló, VAMED,
Nutricia/Danone and Aimmune. TE reports lecture fees from Novartis,
ThermoFisher, Nutricia/Danone, Aimmune, ALK-Abelló.
Acknowledgements: This work was supported by The Canadian
Institutes of Health Research Fredrick Banting and Charles Best Canada
Graduate Scholarship and The Hospital for Sick Children (SickKids)
grants: HSBC Catalyst Grant, POS Innovation Grant, and Restracomp
Graduate Scholarship, as well as the SickKids Food Allergy and
Anaphylaxis Program and start-up funds by SickKids Research Institute
and the Department of Paediatrics.
Abstract (249/250 words):
Background: Food allergy affects up to 8% of the pediatric
population. Despite ongoing efforts, treatment options remain limited.
Novel models of food allergy are needed to study response patterns
downstream of IgE-crosslinking and evaluate drugs modifying acute
events. Here, we report a novel human ex vivo model that displays
acute, allergen-specific, IgE-mediated smooth muscle contractions using
precision cut intestinal slices (PCIS).
Methods: PCIS were generated using gut tissue samples from
children who underwent clinically indicated surgery. Viability and
metabolic activity were assessed from 0-24h. Distribution of relevant
cell subsets was confirmed using single cell nuclear sequencing. PCIS
were passively sensitized using plasma from peanut allergic donors or
peanut-sensitized non-allergic donors, and exposed to various stimuli
including serotonin, histamine, FcɛRI-crosslinker and food allergens.
Smooth muscle contractions and mediator release functioned as readouts.
A novel program designed to measure contractions was developed to
quantify responses. The ability to demonstrate the impact of
antihistamines and immunomodulation from peanut oral immunotherapy (OIT)
was assessed.
Results: PCIS viability was maintained for 24h. Cellular
distribution confirmed the presence of key cell subsets including mast
cells. The video analysis tool reliably quantified responses to
different stimulatory conditions. Smooth muscle contractions were
allergen-specific and reflected the clinical phenotype of the plasma
donor. Tryptase measurement confirmed IgE-dependent mast cell-derived
mediator release. Antihistamines suppressed histamine-induced
contraction and plasma from successful peanut OIT suppressed
peanut-specific PCIS contraction.
Conclusion: PCIS represent a novel human tissue-based model to
study acute, IgE-mediated food allergy and pharmaceutical impacts on
allergic responses in the gut.
Key Words: Allergy model; food allergy; intestine; Precision
Cut Intestinal Slices; PCIS