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Table 3 Methods addressing several MIE’s and KE’s shown in Fig. 1

From: COST Action ‘ImpARAS’: what have we learnt to improve food allergy risk assessment. A summary of a 4 year networking consortium

EventIn vitro methodRead-outs
MIE 1, 2, 3 & KE1M cellsAllergen quantification (SDS-PAGE, Western blot and microscopy)
Integrity of ZO-1 (microscopy).
T84Monolayer integrity (TEER) Cytokine production (ELISA)
HCT-8Monolayer integrity (TEER) Cytokine production (ELISA)
Caco-2Monolayer integrity (TEER and Lucifer Yellow)
Allergen quantification (ELISA, SDS-Page, Western blot and LC–MS,)
Integrity of A20 (Western blot and RT-qPCR blot and microscopy)
Allergen transport (RBL activation test)
Gene expression (RT-qPCR)
HT-29Monolayer integrity (TEER).
Allergen quantification (ELISA)
Integrity of A20 (Western blot and RT-qPCR
KE 2 & 3Mouse BM-DCsAllergen uptake (flow cytometry)
Migration assay (flow cytometry)
Cytokine production (ELISA)
DCs maturation (flow cytometry)
THP-1-derived DCsAllergen uptake (flow cytometry)
Gene expression (RT-qPCR)
Cytokine production (ELISA)
Human Mo-DCsExpression of DC markers (flow cytometry)
KE 4Human T cell clonesT cell proliferation ([3H]-thymidine)
Cytokine production (ELISA)
T cell activation (flow cytometry)
Human peripheral blood mononuclear cells (PBMCs)T cell proliferation (CFSE or [3H]-thymidine)
Cytokine production (flow cytometry)
T cell activation (flow cytometry)
Gene expression (RT-qPCR)
Expression of T cell markers (flow cytometry)
Mouse MLN-isolated T cellsCytokine production (ELISA)
Mouse LP-isolated mononuclear cellsT cell proliferation (CFSE)
Cytokine production (flow cytometry)
KE 2, 3 & 4Co-culture: BM-DCs/primed T cellsAllergen uptake (flow cytometry)
T cell cytokine production (ELISA)
DCs maturation (flow cytometry)