Activation of epithelial cells by the major kiwifruit allergen Act d 1 in human and mouse-derived intestinal model

Nešić, Andrijana; Stam, Annemarie; Čavić, Milena; Klooster, Jean Paul ten; Pieters, Raymond; Smit, Joost; Gavrović-Jankulović, Marija

doi:10.1016/j.jff.2019.103556

Cited by 9 publications

(9 citation statements)

References 37 publications

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“…The cells were allowed to grow for three days to reach 80-90% confluence. The cell monolayers were treated either with activated Act d 1 (1 mg/mL) or with E-64-inactivated Act d 1 (1 mg/mL) as previously described [21]. Act d 1 samples were applied onto the apical side of the cell monolayer, and TEER was measured by using a Millicell ERS meter (Millipore, Bedford, MA, USA).…”

Section: Isolation Of Rna and Qrt-pcr Of Mouse Intestinal Samplesmentioning

confidence: 99%

“…Before the Act d 1 treatment, cells were washed with PBS and incubated in serum-free medium for 1 h. Cells were treated with activated or inactivated Act d 1 (1 mg/mL) for 6 h. Thereafter, the medium was removed and cells were washed with cold PBS and collected by centrifugation (800 rpm, 7 min). Isolation of RNA and evaluation of gene expression were performed as previously described [21]. GAPDH was used as a housekeeping gene to allow normalization of mRNA levels between samples.…”

Section: Isolation Of Rna and Qrt-pcr Of Mouse Intestinal Samplesmentioning

confidence: 99%

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The Kiwifruit Allergen Act d 1 Activates NF-κB Signaling and Affects mRNA Expression of TJ Proteins and Innate Pro-Allergenic Cytokines

et al. 2019

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Impairment of the intestinal barrier is one of the key events in the initiation of the sensitization process in food allergy. The aim of this study was to explore the effects of kiwifruit allergen Act d 1 on intestinal permeability and tight junction protein (TJP) gene expression in vivo and to explore its potential to activate the NF-ĸB signaling pathway and to regulate expression of epithelial pro-allergenic cytokines. Influences of Act d 1 on TJP gene expression and pro-allergenic cytokines in the mouse intestine was analyzed by qPCR upon allergen administration by oral gavage. The effect on the in vivo intestinal permeability was assessed in ELISA by measuring the translocation of β-lactoglobulin (BLG) into circulation. The capacity of Act d 1 to activate the NF-ĸB pathway was tested in HEK293 cells by fluorescent microscopy and flow cytometry. Administration of Actinidin (Act d 1) increased intestinal permeability to the BLG. This was accompanied by changes in gene expression of TJP mRNA and pro-allergenic cytokines IL-25, IL-33, and thymic stromal lymphopoietin (TSLP) compared to the control. Act d 1 reduced TEER of the HEK293 monolayer, was positive in an NF-ĸB-reporter HEK293 cell assay, and induced secretion of TSLP. These findings shed more light on the molecular events in the sensitization process of kiwifruit but possibly also of other protease food allergens.

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Section: Isolation Of Rna and Qrt-pcr Of Mouse Intestinal Samplesmentioning

confidence: 99%

Section: Isolation Of Rna and Qrt-pcr Of Mouse Intestinal Samplesmentioning

confidence: 99%

The Kiwifruit Allergen Act d 1 Activates NF-κB Signaling and Affects mRNA Expression of TJ Proteins and Innate Pro-Allergenic Cytokines

et al. 2019

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“…In order to explore cellular or molecular changes of the cell monolayer posed by allergens or other components suspected to impact on the intestinal integrity, immunohistochemistry, immunofluorescence, Western blotting or gene expression analyses can be performed (Drago et al, 2006;Grozdanovic et al, 2016;Nešić et al, 2019b;Price et al, 2014).…”

Section: In Vitro Methodsmentioning

confidence: 99%

“…In a similar way, intestinal permeability has been assessed in animals with determination of the transport of the cow's milk allergen BLG in different intestinal compartments or in serum using ELISAs (Ballegaard et al, 2021;Graversen et al, 2020b;Nešić et al, 2019b), and has been used to investigate the effects of processing and other food components on the transport of BLG. On a more molecular level, the effect of allergens on barrier integrity can be examined by gene expression or Western blotting analyses for assessment of cell junctional complex proteins (González-González et al, 2019;Nešić et al, 2019a), and morphological changes may be investigated by histology and immunohistochemistry (Ballegaard et al, 2021;Mazzon et al, 2002).…”

Section: Animal Studiesmentioning

confidence: 99%

Intestinal protein uptake and IgE-mediated food allergy

Ballegaard¹,

Bøgh²

2023

Food Research International

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“…It is interesting that the plant-derived food cysteine protease, actinidase, increased intestinal permeability in mice and an in vitro human Caco-2 cell culture model by proteolytical degradation of the key epithelial tight junction transmembrane protein, occludin [ 80 ]. Furthermore, this protease induced release of pro-inflammatory cytokines interleukin (IL)-1β, tumor necrosis factor (TNF) α, and IL-33 in mouse-derived intestinal organoids [ 81 ]. Previous findings indicated that food allergens are involved in the sensitization process in food allergy pathogenesis by challenging enterocytes and compromising the epithelial barrier.…”

Section: Overview Of Morphological Organization Of the Intestinementioning

confidence: 99%

Enterocytes in Food Hypersensitivity Reactions

Ristivojević

Apostolović

Smiljanić

2021

Animals

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Food hypersensitivity reactions are adverse reactions to harmless dietary substances, whose causes are hidden within derangements of the complex immune machinery of humans and mammals. Until recently, enterocytes were considered as solely absorptive cells providing a physical barrier for unwanted lumen constituents. This review focuses on the enterocytes, which are the hub for innate and adaptive immune reactions. Furthermore, the ambiguous nature of enterocytes is also reflected in the fact that enterocytes can be considered as antigen-presenting cells since they constitutively express major histocompatibility complex (MHC) class II molecules. Taken together, it becomes clear that enterocytes have an immense role in maintaining oral tolerance to foreign antigens. In general, the immune system and its mechanisms underlying food hypersensitivity are still unknown and the involvement of components belonging to other anatomical systems, such as enterocytes, in these mechanisms make their elucidation even more difficult. The findings from studies with animal models provide us with valuable information about allergic mechanisms in the animal world, while on the other hand, these models are used to extrapolate results to the pathological conditions occurring in humans. There is a constant need for studies that deal with this topic and can overcome the glitches related to ethics in working with animals.

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Activation of epithelial cells by the major kiwifruit allergen Act d 1 in human and mouse-derived intestinal model

Cited by 9 publications

References 37 publications

The Kiwifruit Allergen Act d 1 Activates NF-κB Signaling and Affects mRNA Expression of TJ Proteins and Innate Pro-Allergenic Cytokines

The Kiwifruit Allergen Act d 1 Activates NF-κB Signaling and Affects mRNA Expression of TJ Proteins and Innate Pro-Allergenic Cytokines

Intestinal protein uptake and IgE-mediated food allergy

Enterocytes in Food Hypersensitivity Reactions

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