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

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Author: Janssen-Duijghuijsen, L.M. · Wichers, H.J. · Norren, K. van · Keijer, J. · Baumert, J.L. · Jong, G.A.H. de · Witkamp, R.F. · Koppelman, S.J.
Type:article
Date:2017
Source:Journal of Immunological Methods, 440, 52–57
Identifier: 575334
Keywords: Nutrition · Ara h 6 · Peanut allergen · Immunoglobulin G · ELISA · Inhibition · Food and Nutrition · Healthy Living · Life · FI - Functional Ingredients · ELSS - Earth, Life and Social Sciences

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. We conclude 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.