JanssenDuijghuijsen Lonneke M, Wichers Harry J, van Norren Klaske, Keijer Jaap, Baumert Joseph L, de Jong Govardus A H, Witkamp Renger F, Koppelman Stef J
Wageningen Food and Biobased Research, Wageningen University and Research, The Netherlands; Nutrition and Pharmacology, Wageningen University and Reserach, The Netherlands; Human and Animal Physiology, Wageningen University and Research, The Netherlands.
Wageningen Food and Biobased Research, Wageningen University and Research, The Netherlands.
J Immunol Methods. 2017 Jan;440:52-57. doi: 10.1016/j.jim.2016.11.002. Epub 2016 Nov 11.
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 100g of light-roasted peanuts. Serum samples were taken at regular intervals for six hours. A double monoclonal sandwich ELISA was 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 that was digested in vitro was 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 interferes with 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 consideration when interpreting immunochemical studies on the absorption of food allergens in serum.
一些研究表明,摄入致敏食物来源后,过敏原可能会出现在循环系统中。然而,报告的血清中过敏原水平较低,且不同研究之间的结论存在差异。在此,我们研究了食用花生后决定血清中过敏原检测的因素。10名健康志愿者食用了100克轻度烤制的花生。在6小时内定期采集血清样本。采用双单克隆夹心ELISA法分析血清中主要花生过敏原Ara h 6的存在情况和数量。10名受试者中有4名未检测到Ara h 6。体外消化的纯化Ara h 6在ELISA中仍具有反应性,排除了消化产生无法检测到的小肽的可能性。在基线血清中加入纯化的Ara h 6表明,这4名受试者的摄入前血清部分阻止了Ara h 6在ELISA中的反应,反应性降低高达3个数量级或更多。其他6名受试者的摄入前血清未显示出这种效果。Ara h 6反应性的降低与高滴度的IgG和IgG4同时出现,从摄入前血清中去除IgG可完全消除这种效果,表明IgG和IgG4抑制了Ara h6在ELISA中的反应性。我们得出结论,一些个体血液中存在针对食物过敏原的IgG和IgG4,这会干扰血清中此类食物过敏原的检测。由于这种效应并非在每个个体中都出现,因此在解释关于血清中食物过敏原吸收的免疫化学研究结果时,应考虑到这种干扰的可能性。