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European Journal of Nutrition

, Volume 58, Issue 4, pp 1507–1514 | Cite as

Dietary wheat amylase trypsin inhibitors exacerbate murine allergic airway inflammation

  • Victor F. ZevallosEmail author
  • Verena K. Raker
  • Joachim Maxeiner
  • Petra Scholtes
  • Kerstin Steinbrink
  • Detlef Schuppan
Original Contribution

Abstract

Background

Wheat amylase trypsin inhibitors (ATI) are dietary non-gluten proteins that activate the toll-like receptor 4 on myeloid cells, promoting intestinal inflammation.

Aim of the study

We investigated the effects of dietary ATI on experimental allergic airway inflammation.

Methods

Mice on a gluten and ATI-free diet (GAFD), sensitized with PBS or ovalbumin (OVA) and challenged with OVA, were compared to mice on a commercial standard chow, a gluten diet naturally containing ~ 0.75% of protein as ATI (G+AD), a gluten diet containing ~ 0.19% of protein as ATI (G−AD) and a GAFD with 1% of protein as ATI (AD). Airway hyperreactivity (AHR), inflammation in bronchoalveolar lavage (BAL) and pulmonary tissue sections were analyzed. Allergic sensitization was assessed ex vivo via proliferation of OVA-stimulated splenocytes.

Results

Mice on a GAFD sensitized with PBS did not develop AHR after local provocation with methacholine. Mice on a GAFD or on a G−AD and sensitized with OVA developed milder AHR compared to mice fed a G+AD or an AD. The increased AHR was paralleled by increased BAL eosinophils, IL-5 and IL-13 production, and an enhanced ex vivo splenocyte activation in the ATI-fed groups.

Conclusions

Dietary ATI enhance allergic airway inflammation in OVA-challenged mice, while an ATI-free or ATI-reduced diet has a protective effect on AHR. Nutritional wheat ATI, activators of intestinal myeloid cells, may be clinically relevant adjuvants to allergic airway inflammation.

Keywords

Amylase trypsin inhibitors Gluten Wheat sensitivity Innate immunity Allergic airway inflammation 

Abbreviations

AAI

Allergic airway inflammation

AHR

Airway hyper-reactivity

ATI

Amylase trypsin inhibitor

BAL

Bronchoalveolar lavage

G

Gluten

GFD

Gluten-free diet

HLA

Human lymphocyte antigen

IL

Interleukin

NCWS

Non-celiac (non-allergy) wheat sensitivity

OVA

Ovalbumin

PAS

Periodic acid–Schiff

TLR

Toll-like receptor

Notes

Acknowledgements

This study was supported by Grants of the German Research Foundation (DFG Schu646/17-1) and by the Leibniz-Foundation (Project WheatScan).

Author contributions

VZ: study design; acquisition of data; analysis and interpretation of data; drafting and editing of the manuscript; statistical analysis. VR: acquisition of data and support during experiment and data acquisition/analysis (stimulation of splenocytes). JM and PS: acquisition of data and support during in-vivo experiments (lung measurements). KS and other authors: support in revising the manuscript. DS: study design; interpretation of data; study supervision; drafting and editing of the manuscript.

Compliance with ethical standards

Conflict of interest

On behalf of all authors, the corresponding author states that there is no conflict of interest.

Supplementary material

394_2018_1681_MOESM1_ESM.jpg (737 kb)
Supplementary Figure 1. Standard chow diet contains biological active ATI. (A) Experimental protocol, C57BL/6 mice (n=6) were raised on a standard chow (SC) diet, continued on this diet or placed on a defined GAFD for four weeks and then divided in 4 groups: group 1 and 2, continued on SC, and either mock-sensitised three times with PBS or sensitized with OVA; group 3 and 4, were placed on a GAFD, mock-sensitized with PBS or sensitized with OVA. All groups were challenged with OVA. (B) Differential cell counts in BAL. Mac, macrophages; neut, neutrophils; eos, eosinophils; lymph, lymphocytes. (C) Representative H&E-stained lung tissue from all groups. Scale bars, 200 μm. (D) H&E score of all groups (E) IL-13 levels in BAL. All results are representative of two different experiments and are expressed as the mean ± SEM (n=6). Statistical significance was determined with two-way analysis of variance (ANOVA) (JPG 737 KB)
394_2018_1681_MOESM2_ESM.jpg (690 kb)
Supplementary figure 2. Standard chow diet contains biological active ATI. (A) Representative Periodic acid–Schiff (PAS)-stained lung tissue from all groups of mice. Scale bars, 200 μm. (B) PAS score of all groups. (C) Measurement of AHR, assessed by airway resistance to increasing doses of methacholine. (D) TLR4-stimulating bioactivity in protein extracts from SC and GAFD. All results are representative of data of two different experiments and are expressed as the mean ± SEM (n=6). Statistical significance was determined with two-way analysis of variance (ANOVA). ***p< 0.001 (JPG 690 KB)
394_2018_1681_MOESM3_ESM.docx (14 kb)
Supplementary material 3 (DOCX 13 KB)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Victor F. Zevallos
    • 1
    • 2
    Email author
  • Verena K. Raker
    • 2
    • 3
  • Joachim Maxeiner
    • 2
    • 4
  • Petra Scholtes
    • 2
    • 4
  • Kerstin Steinbrink
    • 2
    • 3
  • Detlef Schuppan
    • 1
    • 2
    • 5
  1. 1.Institute of Translational Immunology, University Medical CenterJohannes Gutenberg University MainzMainzGermany
  2. 2.Research Center for Immunotherapy, University Medical CenterJohannes Gutenberg University MainzMainzGermany
  3. 3.Department of Dermatology, University Medical CenterJohannes Gutenberg University MainzMainzGermany
  4. 4.Asthma Core Facility, University Medical CenterJohannes Gutenberg University MainzMainzGermany
  5. 5.Division of Gastroenterology, Beth Israel Deaconess Medical CenterHarvard Medical SchoolBostonUSA

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