Anisakis simplex products impair intestinal epithelial barrier function and occludin and zonula occludens-1 localisation in differentiated Caco-2 cells

[Background]: Anisakis spp. are nematode parasites found in a wide range of marine organisms. Human beings may accidentally become infected, showing the symptoms of anisakiasis and allergic responses. There has been evidence of increased intestinal permeability in A. simplex¿sensitized subjects and that specific IgE titres increase in some allergic patients when fishery products are re-introduced into their diet. The aims of this work were to study the effect of A. simplex crude extract on the intestinal integrity and permeability by using Caco-2 cell monolayer. To analyse the capacity of Ani s 4 allergen to cross the epithelial barrier. [Methodology/Principal findings]: Cellular bioenergetics, transepithelial electrical resistance, viability, permeability, reactive oxygen species generation and immunofluorescent staining of tight junction proteins were analysed. A. simplex crude extract compromises the Caco-2 cell monolayer integrity in a dose-dependent manner. It is a rapid and recoverable effect. The epithelial barrier disruption is accompanied by an increase in paracellular permeability and reactive oxygen species production and by a delocalization of occludin and zonula occludens-1. Finally, the allergen Ani s 4 is able to cross the epithelial barrier in Caco-2 monolayer and reach a cumulative mean percentage of 22.7% of total concentration in the basolateral side after 24 hours of culture. [Conclusions/Significance]: Our results demonstrate that A. simplex induces an early and reversible alteration of integrity and permeability of Caco-2 cell monolayer and that an underlying mechanism of this effect would involve the oxidative stress and disruption of epithelial tight junctions. Additionally, it has been shown that Ani s 4 allergen is able to cross the epithelial barrier. These findings could explain the increased intestinal permeability observed in Anisakis-sensitized patients, the changes over time in IgE sensitization to A. simplex allergens, and the specific IgE persistence in Anisakis allergy.

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Main Authors: Carballeda-Sangiao, Noelia, Sánchez-Alonso, Isabel, Navas, Alfonso, Arcos, Susana C., Fernández de Palencia, P., Careche, Mercedes, González Muñoz, Miguel
Other Authors: Ministerio de Economía y Competitividad (España)
Format: artículo biblioteca
Published: 2020-07-06
Online Access:http://hdl.handle.net/10261/235351
http://dx.doi.org/10.13039/501100000780
http://dx.doi.org/10.13039/501100003329
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spelling dig-ictan-es-10261-2353512022-01-03T12:08:02Z Anisakis simplex products impair intestinal epithelial barrier function and occludin and zonula occludens-1 localisation in differentiated Caco-2 cells Carballeda-Sangiao, Noelia Sánchez-Alonso, Isabel Navas, Alfonso Arcos, Susana C. Fernández de Palencia, P. Careche, Mercedes González Muñoz, Miguel Ministerio de Economía y Competitividad (España) European Commission [Background]: Anisakis spp. are nematode parasites found in a wide range of marine organisms. Human beings may accidentally become infected, showing the symptoms of anisakiasis and allergic responses. There has been evidence of increased intestinal permeability in A. simplex¿sensitized subjects and that specific IgE titres increase in some allergic patients when fishery products are re-introduced into their diet. The aims of this work were to study the effect of A. simplex crude extract on the intestinal integrity and permeability by using Caco-2 cell monolayer. To analyse the capacity of Ani s 4 allergen to cross the epithelial barrier. [Methodology/Principal findings]: Cellular bioenergetics, transepithelial electrical resistance, viability, permeability, reactive oxygen species generation and immunofluorescent staining of tight junction proteins were analysed. A. simplex crude extract compromises the Caco-2 cell monolayer integrity in a dose-dependent manner. It is a rapid and recoverable effect. The epithelial barrier disruption is accompanied by an increase in paracellular permeability and reactive oxygen species production and by a delocalization of occludin and zonula occludens-1. Finally, the allergen Ani s 4 is able to cross the epithelial barrier in Caco-2 monolayer and reach a cumulative mean percentage of 22.7% of total concentration in the basolateral side after 24 hours of culture. [Conclusions/Significance]: Our results demonstrate that A. simplex induces an early and reversible alteration of integrity and permeability of Caco-2 cell monolayer and that an underlying mechanism of this effect would involve the oxidative stress and disruption of epithelial tight junctions. Additionally, it has been shown that Ani s 4 allergen is able to cross the epithelial barrier. These findings could explain the increased intestinal permeability observed in Anisakis-sensitized patients, the changes over time in IgE sensitization to A. simplex allergens, and the specific IgE persistence in Anisakis allergy. The research leading to these results was supported by the Project ANIRISK (AGL2015-68248-C2) under the Research, Development and Innovation Programme oriented to Societal Challenges funded by the Spanish Ministry of Economy and Competitiveness (MINECO) and European Regional Development Fund (FEDER). 2021-03-22T09:00:28Z 2021-03-22T09:00:28Z 2020-07-06 2021-03-22T09:00:29Z artículo http://purl.org/coar/resource_type/c_6501 doi: 10.1371/journal.pntd.0008462 issn: 1935-2727 PLoS Neglected Tropical Diseases 14(7): e0008462 (2020) http://hdl.handle.net/10261/235351 10.1371/journal.pntd.0008462 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100003329 #PLACEHOLDER_PARENT_METADATA_VALUE# info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/AGL2015-68248-C2 Publisher's version http://dx.doi.org/10.1371/journal.pntd.0008462 Sí open
institution ICTAN ES
collection DSpace
country España
countrycode ES
component Bibliográfico
access En linea
databasecode dig-ictan-es
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region Europa del Sur
libraryname Biblioteca del ICTAN España
description [Background]: Anisakis spp. are nematode parasites found in a wide range of marine organisms. Human beings may accidentally become infected, showing the symptoms of anisakiasis and allergic responses. There has been evidence of increased intestinal permeability in A. simplex¿sensitized subjects and that specific IgE titres increase in some allergic patients when fishery products are re-introduced into their diet. The aims of this work were to study the effect of A. simplex crude extract on the intestinal integrity and permeability by using Caco-2 cell monolayer. To analyse the capacity of Ani s 4 allergen to cross the epithelial barrier. [Methodology/Principal findings]: Cellular bioenergetics, transepithelial electrical resistance, viability, permeability, reactive oxygen species generation and immunofluorescent staining of tight junction proteins were analysed. A. simplex crude extract compromises the Caco-2 cell monolayer integrity in a dose-dependent manner. It is a rapid and recoverable effect. The epithelial barrier disruption is accompanied by an increase in paracellular permeability and reactive oxygen species production and by a delocalization of occludin and zonula occludens-1. Finally, the allergen Ani s 4 is able to cross the epithelial barrier in Caco-2 monolayer and reach a cumulative mean percentage of 22.7% of total concentration in the basolateral side after 24 hours of culture. [Conclusions/Significance]: Our results demonstrate that A. simplex induces an early and reversible alteration of integrity and permeability of Caco-2 cell monolayer and that an underlying mechanism of this effect would involve the oxidative stress and disruption of epithelial tight junctions. Additionally, it has been shown that Ani s 4 allergen is able to cross the epithelial barrier. These findings could explain the increased intestinal permeability observed in Anisakis-sensitized patients, the changes over time in IgE sensitization to A. simplex allergens, and the specific IgE persistence in Anisakis allergy.
author2 Ministerio de Economía y Competitividad (España)
author_facet Ministerio de Economía y Competitividad (España)
Carballeda-Sangiao, Noelia
Sánchez-Alonso, Isabel
Navas, Alfonso
Arcos, Susana C.
Fernández de Palencia, P.
Careche, Mercedes
González Muñoz, Miguel
format artículo
author Carballeda-Sangiao, Noelia
Sánchez-Alonso, Isabel
Navas, Alfonso
Arcos, Susana C.
Fernández de Palencia, P.
Careche, Mercedes
González Muñoz, Miguel
spellingShingle Carballeda-Sangiao, Noelia
Sánchez-Alonso, Isabel
Navas, Alfonso
Arcos, Susana C.
Fernández de Palencia, P.
Careche, Mercedes
González Muñoz, Miguel
Anisakis simplex products impair intestinal epithelial barrier function and occludin and zonula occludens-1 localisation in differentiated Caco-2 cells
author_sort Carballeda-Sangiao, Noelia
title Anisakis simplex products impair intestinal epithelial barrier function and occludin and zonula occludens-1 localisation in differentiated Caco-2 cells
title_short Anisakis simplex products impair intestinal epithelial barrier function and occludin and zonula occludens-1 localisation in differentiated Caco-2 cells
title_full Anisakis simplex products impair intestinal epithelial barrier function and occludin and zonula occludens-1 localisation in differentiated Caco-2 cells
title_fullStr Anisakis simplex products impair intestinal epithelial barrier function and occludin and zonula occludens-1 localisation in differentiated Caco-2 cells
title_full_unstemmed Anisakis simplex products impair intestinal epithelial barrier function and occludin and zonula occludens-1 localisation in differentiated Caco-2 cells
title_sort anisakis simplex products impair intestinal epithelial barrier function and occludin and zonula occludens-1 localisation in differentiated caco-2 cells
publishDate 2020-07-06
url http://hdl.handle.net/10261/235351
http://dx.doi.org/10.13039/501100000780
http://dx.doi.org/10.13039/501100003329
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