Detection of peanut allergen in human blood after consumption of peanuts is skewed by endogenous immunoglobulins

L.M. Janssen Duijghuijsen, H.J. Wichers, K. van Norren, J. Keijer, J.L. Baumert, Govardus A.H. De Jong, R.F. Witkamp, S.J. Koppelman

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Abstract

Some studies have suggested that allergens may appear in the circulation after ingestion of allergenic food
sources. The reported levels of allergen in serum, however, are low, and conclusions between studies differ.
Here, we investigated factors that determine the detection of allergens in serum after consumption of peanuts.
Ten healthy volunteers ingested 100 g of light-roasted peanuts. Serumsampleswere taken at regular intervals for
six hours. A double monoclonal sandwich ELISAwas used to analyse the presence and quantity of the major peanut
allergen Ara h 6 in serum.
In 4 out of 10 subjects, no Ara h 6 could be detected. Purified Ara h 6 thatwas digested in vitrowas still reactive in
the ELISA, rejecting the possibility that digestion leads to small peptides that could not be detected. Spiking of purified
Ara h 6 in baseline serum showed that the pre-ingestion serum of these four subjects partially prevented
Ara h 6 to react in the ELISA, with a reduction of reactivity of up to 3 orders of magnitude or more. Pre-ingestion
serum of the other six subjects did not show such an effect. The reduction of reactivity of Ara h 6 coincided with
high titres of IgG and IgG4, and removal of IgG from pre-ingestion serum abolished this effect completely, indicating
that IgG and IgG4 inhibited the reactivity of Ara h6 in the ELISA.
Weconclude that some individuals have IgG and IgG4 against food allergens in their blood, which interfereswith
detection of such food allergens in serum. Because this effect does not occur for each individual, the possibility of
such interference should be taken into considerationwhen interpreting immunochemical studies on the absorption
of food allergens in serum.
LanguageEnglish
Pages52-57
JournalJournal of Immunological Methods
Volume440
DOIs
Publication statusPublished - 2017

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Allergens
Immunoglobulins
Immunoglobulin G
Serum
Eating
Enzyme-Linked Immunosorbent Assay
Food
Arachis
Digestion
Healthy Volunteers
Light
Peptides

Cite this

@article{ac45f9754e594b19831259ddfdabc4d7,
title = "Detection of peanut allergen in human blood after consumption of peanuts is skewed by endogenous immunoglobulins",
abstract = "Some studies have suggested that allergens may appear in the circulation after ingestion of allergenic foodsources. The reported levels of allergen in serum, however, are low, and conclusions between studies differ.Here, we investigated factors that determine the detection of allergens in serum after consumption of peanuts.Ten healthy volunteers ingested 100 g of light-roasted peanuts. Serumsampleswere taken at regular intervals forsix hours. A double monoclonal sandwich ELISAwas used to analyse the presence and quantity of the major peanutallergen Ara h 6 in serum.In 4 out of 10 subjects, no Ara h 6 could be detected. Purified Ara h 6 thatwas digested in vitrowas still reactive inthe ELISA, rejecting the possibility that digestion leads to small peptides that could not be detected. Spiking of purifiedAra h 6 in baseline serum showed that the pre-ingestion serum of these four subjects partially preventedAra h 6 to react in the ELISA, with a reduction of reactivity of up to 3 orders of magnitude or more. Pre-ingestionserum of the other six subjects did not show such an effect. The reduction of reactivity of Ara h 6 coincided withhigh titres of IgG and IgG4, and removal of IgG from pre-ingestion serum abolished this effect completely, indicatingthat IgG and IgG4 inhibited the reactivity of Ara h6 in the ELISA.Weconclude that some individuals have IgG and IgG4 against food allergens in their blood, which interfereswithdetection of such food allergens in serum. Because this effect does not occur for each individual, the possibility ofsuch interference should be taken into considerationwhen interpreting immunochemical studies on the absorptionof food allergens in serum.",
author = "{Janssen Duijghuijsen}, L.M. and H.J. Wichers and {van Norren}, K. and J. Keijer and J.L. Baumert and {De Jong}, {Govardus A.H.} and R.F. Witkamp and S.J. Koppelman",
year = "2017",
doi = "10.1016/j.jim.2016.11.002",
language = "English",
volume = "440",
pages = "52--57",
journal = "Journal of Immunological Methods",
issn = "0022-1759",
publisher = "Elsevier",

}

Detection of peanut allergen in human blood after consumption of peanuts is skewed by endogenous immunoglobulins. / Janssen Duijghuijsen, L.M.; Wichers, H.J.; van Norren, K.; Keijer, J.; Baumert, J.L.; De Jong, Govardus A.H.; Witkamp, R.F.; Koppelman, S.J.

In: Journal of Immunological Methods, Vol. 440, 2017, p. 52-57.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Detection of peanut allergen in human blood after consumption of peanuts is skewed by endogenous immunoglobulins

AU - Janssen Duijghuijsen, L.M.

AU - Wichers, H.J.

AU - van Norren, K.

AU - Keijer, J.

AU - Baumert, J.L.

AU - De Jong, Govardus A.H.

AU - Witkamp, R.F.

AU - Koppelman, S.J.

PY - 2017

Y1 - 2017

N2 - Some studies have suggested that allergens may appear in the circulation after ingestion of allergenic foodsources. The reported levels of allergen in serum, however, are low, and conclusions between studies differ.Here, we investigated factors that determine the detection of allergens in serum after consumption of peanuts.Ten healthy volunteers ingested 100 g of light-roasted peanuts. Serumsampleswere taken at regular intervals forsix hours. A double monoclonal sandwich ELISAwas used to analyse the presence and quantity of the major peanutallergen Ara h 6 in serum.In 4 out of 10 subjects, no Ara h 6 could be detected. Purified Ara h 6 thatwas digested in vitrowas still reactive inthe ELISA, rejecting the possibility that digestion leads to small peptides that could not be detected. Spiking of purifiedAra h 6 in baseline serum showed that the pre-ingestion serum of these four subjects partially preventedAra h 6 to react in the ELISA, with a reduction of reactivity of up to 3 orders of magnitude or more. Pre-ingestionserum of the other six subjects did not show such an effect. The reduction of reactivity of Ara h 6 coincided withhigh titres of IgG and IgG4, and removal of IgG from pre-ingestion serum abolished this effect completely, indicatingthat IgG and IgG4 inhibited the reactivity of Ara h6 in the ELISA.Weconclude that some individuals have IgG and IgG4 against food allergens in their blood, which interfereswithdetection of such food allergens in serum. Because this effect does not occur for each individual, the possibility ofsuch interference should be taken into considerationwhen interpreting immunochemical studies on the absorptionof food allergens in serum.

AB - Some studies have suggested that allergens may appear in the circulation after ingestion of allergenic foodsources. The reported levels of allergen in serum, however, are low, and conclusions between studies differ.Here, we investigated factors that determine the detection of allergens in serum after consumption of peanuts.Ten healthy volunteers ingested 100 g of light-roasted peanuts. Serumsampleswere taken at regular intervals forsix hours. A double monoclonal sandwich ELISAwas used to analyse the presence and quantity of the major peanutallergen Ara h 6 in serum.In 4 out of 10 subjects, no Ara h 6 could be detected. Purified Ara h 6 thatwas digested in vitrowas still reactive inthe ELISA, rejecting the possibility that digestion leads to small peptides that could not be detected. Spiking of purifiedAra h 6 in baseline serum showed that the pre-ingestion serum of these four subjects partially preventedAra h 6 to react in the ELISA, with a reduction of reactivity of up to 3 orders of magnitude or more. Pre-ingestionserum of the other six subjects did not show such an effect. The reduction of reactivity of Ara h 6 coincided withhigh titres of IgG and IgG4, and removal of IgG from pre-ingestion serum abolished this effect completely, indicatingthat IgG and IgG4 inhibited the reactivity of Ara h6 in the ELISA.Weconclude that some individuals have IgG and IgG4 against food allergens in their blood, which interfereswithdetection of such food allergens in serum. Because this effect does not occur for each individual, the possibility ofsuch interference should be taken into considerationwhen interpreting immunochemical studies on the absorptionof food allergens in serum.

U2 - 10.1016/j.jim.2016.11.002

DO - 10.1016/j.jim.2016.11.002

M3 - Article

VL - 440

SP - 52

EP - 57

JO - Journal of Immunological Methods

T2 - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

ER -