First successful reduction of clinical allergenicity of food by genetic modification: Mal d 1 silenced apples cause fewer allergy symptoms than the wild-type cultivar

A.E.J. Dubois, G. Pagliarani, R.M. Brouwer, B.J. Kollen, L.O. Dragsted, F.D. Eriksen, O. Callesen, L.J.W.J. Gilissen, F.A. Krens, R.G.F. Visser, M.J.M. Smulders, B.J. Vlieg-Boerstra, B.J. Flokstra-de Blok, W.E. van de Weg

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Abstract

Background Genetic modification of allergenic foods such as apple has the potential to reduce their clinical allergenicity, but this has never been studied by oral challenges in allergic individuals. Methods We performed oral food challenges in 21 apple-allergic individuals with Elstar apples which had undergone gene silencing of the major allergen of apple, Mal d 1, by RNA interference. Downregulation of Mal d 1 gene expression in the apples was verified by qRT-PCR. Clinical responses to the genetically modified apples were compared to those seen with the wild-type Elstar using a visual analogue scale (VAS). Results Gene silencing produced two genetically modified apple lines expressing Mal d 1.02 and other Mal d 1 gene mRNA levels which were extensively downregulated, that is only 0.1–16.4% (e-DR1) and 0.2–9.9% (e-DR2) of those of the wild-type Elstar, respectively. Challenges with these downregulated apple lines produced significantly less intense maximal symptoms to the first dose (Vmax1) than with Elstar (Vmax1 Elstar 3.0 mm vs 0.0 mm for e-DR1, P = 0.017 and 0.0 mm for e-DR2, P = 0.043), as well as significantly less intense mean symptoms per dose (meanV/d) than with Elstar (meanV/d Elstar 2.2 mm vs 0.2 mm for e-DR1, P = 0.017 and 0.0 mm for e-DR2, P = 0.043). Only one subject (5%) remained symptom-free when challenged with the Elstar apple, whereas 43% did so with e-DR1 and 63% with e-DR2. Conclusion These data show that mRNA silencing of Mal d 1 results in a marked reduction of Mal d 1 gene expression in the fruit and reduction of symptoms when these apples are ingested by allergic subjects. Approximately half of the subjects developed no symptoms whatsoever, and virtually all subjects wished to consume the apple again in the future.
Original languageEnglish
Pages (from-to)1406-1412
JournalAllergy
Volume70
Issue number11
DOIs
Publication statusPublished - 2015

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Malus
Hypersensitivity
Food
Down-Regulation
Gene Expression
Messenger RNA
Gene Silencing
RNA Interference
Visual Analog Scale
Allergens
Fruit
Polymerase Chain Reaction

Cite this

Dubois, A.E.J. ; Pagliarani, G. ; Brouwer, R.M. ; Kollen, B.J. ; Dragsted, L.O. ; Eriksen, F.D. ; Callesen, O. ; Gilissen, L.J.W.J. ; Krens, F.A. ; Visser, R.G.F. ; Smulders, M.J.M. ; Vlieg-Boerstra, B.J. ; Flokstra-de Blok, B.J. ; van de Weg, W.E. / First successful reduction of clinical allergenicity of food by genetic modification: Mal d 1 silenced apples cause fewer allergy symptoms than the wild-type cultivar. In: Allergy. 2015 ; Vol. 70, No. 11. pp. 1406-1412.
@article{e96c7328e2ce4f3989107dec0c3c4f50,
title = "First successful reduction of clinical allergenicity of food by genetic modification: Mal d 1 silenced apples cause fewer allergy symptoms than the wild-type cultivar",
abstract = "Background Genetic modification of allergenic foods such as apple has the potential to reduce their clinical allergenicity, but this has never been studied by oral challenges in allergic individuals. Methods We performed oral food challenges in 21 apple-allergic individuals with Elstar apples which had undergone gene silencing of the major allergen of apple, Mal d 1, by RNA interference. Downregulation of Mal d 1 gene expression in the apples was verified by qRT-PCR. Clinical responses to the genetically modified apples were compared to those seen with the wild-type Elstar using a visual analogue scale (VAS). Results Gene silencing produced two genetically modified apple lines expressing Mal d 1.02 and other Mal d 1 gene mRNA levels which were extensively downregulated, that is only 0.1–16.4{\%} (e-DR1) and 0.2–9.9{\%} (e-DR2) of those of the wild-type Elstar, respectively. Challenges with these downregulated apple lines produced significantly less intense maximal symptoms to the first dose (Vmax1) than with Elstar (Vmax1 Elstar 3.0 mm vs 0.0 mm for e-DR1, P = 0.017 and 0.0 mm for e-DR2, P = 0.043), as well as significantly less intense mean symptoms per dose (meanV/d) than with Elstar (meanV/d Elstar 2.2 mm vs 0.2 mm for e-DR1, P = 0.017 and 0.0 mm for e-DR2, P = 0.043). Only one subject (5{\%}) remained symptom-free when challenged with the Elstar apple, whereas 43{\%} did so with e-DR1 and 63{\%} with e-DR2. Conclusion These data show that mRNA silencing of Mal d 1 results in a marked reduction of Mal d 1 gene expression in the fruit and reduction of symptoms when these apples are ingested by allergic subjects. Approximately half of the subjects developed no symptoms whatsoever, and virtually all subjects wished to consume the apple again in the future.",
author = "A.E.J. Dubois and G. Pagliarani and R.M. Brouwer and B.J. Kollen and L.O. Dragsted and F.D. Eriksen and O. Callesen and L.J.W.J. Gilissen and F.A. Krens and R.G.F. Visser and M.J.M. Smulders and B.J. Vlieg-Boerstra and {Flokstra-de Blok}, B.J. and {van de Weg}, W.E.",
year = "2015",
doi = "10.1111/all.12684",
language = "English",
volume = "70",
pages = "1406--1412",
journal = "Allergy",
issn = "0105-4538",
publisher = "Wiley",
number = "11",

}

First successful reduction of clinical allergenicity of food by genetic modification: Mal d 1 silenced apples cause fewer allergy symptoms than the wild-type cultivar. / Dubois, A.E.J.; Pagliarani, G.; Brouwer, R.M.; Kollen, B.J.; Dragsted, L.O.; Eriksen, F.D.; Callesen, O.; Gilissen, L.J.W.J.; Krens, F.A.; Visser, R.G.F.; Smulders, M.J.M.; Vlieg-Boerstra, B.J.; Flokstra-de Blok, B.J.; van de Weg, W.E.

In: Allergy, Vol. 70, No. 11, 2015, p. 1406-1412.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - First successful reduction of clinical allergenicity of food by genetic modification: Mal d 1 silenced apples cause fewer allergy symptoms than the wild-type cultivar

AU - Dubois, A.E.J.

AU - Pagliarani, G.

AU - Brouwer, R.M.

AU - Kollen, B.J.

AU - Dragsted, L.O.

AU - Eriksen, F.D.

AU - Callesen, O.

AU - Gilissen, L.J.W.J.

AU - Krens, F.A.

AU - Visser, R.G.F.

AU - Smulders, M.J.M.

AU - Vlieg-Boerstra, B.J.

AU - Flokstra-de Blok, B.J.

AU - van de Weg, W.E.

PY - 2015

Y1 - 2015

N2 - Background Genetic modification of allergenic foods such as apple has the potential to reduce their clinical allergenicity, but this has never been studied by oral challenges in allergic individuals. Methods We performed oral food challenges in 21 apple-allergic individuals with Elstar apples which had undergone gene silencing of the major allergen of apple, Mal d 1, by RNA interference. Downregulation of Mal d 1 gene expression in the apples was verified by qRT-PCR. Clinical responses to the genetically modified apples were compared to those seen with the wild-type Elstar using a visual analogue scale (VAS). Results Gene silencing produced two genetically modified apple lines expressing Mal d 1.02 and other Mal d 1 gene mRNA levels which were extensively downregulated, that is only 0.1–16.4% (e-DR1) and 0.2–9.9% (e-DR2) of those of the wild-type Elstar, respectively. Challenges with these downregulated apple lines produced significantly less intense maximal symptoms to the first dose (Vmax1) than with Elstar (Vmax1 Elstar 3.0 mm vs 0.0 mm for e-DR1, P = 0.017 and 0.0 mm for e-DR2, P = 0.043), as well as significantly less intense mean symptoms per dose (meanV/d) than with Elstar (meanV/d Elstar 2.2 mm vs 0.2 mm for e-DR1, P = 0.017 and 0.0 mm for e-DR2, P = 0.043). Only one subject (5%) remained symptom-free when challenged with the Elstar apple, whereas 43% did so with e-DR1 and 63% with e-DR2. Conclusion These data show that mRNA silencing of Mal d 1 results in a marked reduction of Mal d 1 gene expression in the fruit and reduction of symptoms when these apples are ingested by allergic subjects. Approximately half of the subjects developed no symptoms whatsoever, and virtually all subjects wished to consume the apple again in the future.

AB - Background Genetic modification of allergenic foods such as apple has the potential to reduce their clinical allergenicity, but this has never been studied by oral challenges in allergic individuals. Methods We performed oral food challenges in 21 apple-allergic individuals with Elstar apples which had undergone gene silencing of the major allergen of apple, Mal d 1, by RNA interference. Downregulation of Mal d 1 gene expression in the apples was verified by qRT-PCR. Clinical responses to the genetically modified apples were compared to those seen with the wild-type Elstar using a visual analogue scale (VAS). Results Gene silencing produced two genetically modified apple lines expressing Mal d 1.02 and other Mal d 1 gene mRNA levels which were extensively downregulated, that is only 0.1–16.4% (e-DR1) and 0.2–9.9% (e-DR2) of those of the wild-type Elstar, respectively. Challenges with these downregulated apple lines produced significantly less intense maximal symptoms to the first dose (Vmax1) than with Elstar (Vmax1 Elstar 3.0 mm vs 0.0 mm for e-DR1, P = 0.017 and 0.0 mm for e-DR2, P = 0.043), as well as significantly less intense mean symptoms per dose (meanV/d) than with Elstar (meanV/d Elstar 2.2 mm vs 0.2 mm for e-DR1, P = 0.017 and 0.0 mm for e-DR2, P = 0.043). Only one subject (5%) remained symptom-free when challenged with the Elstar apple, whereas 43% did so with e-DR1 and 63% with e-DR2. Conclusion These data show that mRNA silencing of Mal d 1 results in a marked reduction of Mal d 1 gene expression in the fruit and reduction of symptoms when these apples are ingested by allergic subjects. Approximately half of the subjects developed no symptoms whatsoever, and virtually all subjects wished to consume the apple again in the future.

U2 - 10.1111/all.12684

DO - 10.1111/all.12684

M3 - Article

VL - 70

SP - 1406

EP - 1412

JO - Allergy

JF - Allergy

SN - 0105-4538

IS - 11

ER -