Prospective investigation on the transfer of Ara h 2, the most potent peanut allergen, in human breast milk
Prospective investigation on the transfer of Ara h 2, the most potent peanut allergen, in human breast milk
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DOI：
10.1111/pai.12533
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发表时间：
2016-06-01
影响因子：
4.4
通讯作者：
Jappe, Uta
中科院分区：
文献类型：
--
作者：
Schocker, Frauke;Baumert, Joseph;Jappe, Uta
背景：花生过敏是最严重的食物过敏之一。母乳喂养是否会导致对花生的耐受性，或者相反，是否会预置风险--婴儿对花生的隐匿性过敏反应仍是一个讨论的问题。在德国的一项母乳研究中，我们试图调查最有效的花生过敏原arah2在母乳中的转移情况，并阐明arah2出现的时间动力学。方法：我们招募了32名哺乳期、不对花生过敏的妇女，在饮用100g干烘烤花生后的不同时间点采集母乳样本。采用不同的免疫学方法对母乳样品中的Arah2进行检测：用患者血清进行2D免疫印迹，用抗Arah2的单抗亲和富集法，然后基于LC-MS/MS检测，以及竞争抑制ELISA法检测Arah2及其抗消化多肽(DRP-Arah2)。结果：在定性分析中，利用患者血清的2D免疫印迹和免疫亲和富集法，再经LC-MS/MS分析，可以在母乳样品中检测到Arah2。在半定量分析中，32例受试者中有9例在母乳中检测到Arah2及其耐消化性多肽。有证据表明，阿糖胞苷在母乳中以不同浓度迅速(1、2、3、4h)或延迟(8h、12h)单独排泄。母乳中阿糖胞苷的鉴定是研究其致敏或耐受特性的前提。
Background: Peanut allergy is one of the most severe food allergies. Whether breastfeeding induces tolerance to peanuts or on the contrary, pre-disposes at risk-babies to occult allergic sensitization to peanuts is still a matter of discussion. We sought to investigate the transfer of the most potent peanut allergen Ara h 2 into human breast milk in a German breast milk study and to shed light on the time kinetics of Ara h 2 appearance.Methods: We recruited 32 lactating, non-peanut-allergic women and collected breast milk samples at different time points after consumption of 100 g dry roasted peanuts. Breast milk samples were investigated for Ara h 2 with different immunological methods: by 2D immunoblotting with a patient's serum, by affinity enrichment using a monoclonal antibody against Ara h 2 followed by LC-MS/MS-based detection and by a competitive inhibition ELISA for the detection of Ara h 2 and its digestion-resistant peptides (DRP-Ara h 2).Results: In a qualitative analysis, Ara h 2 could be identified in a breast milk sample by 2D immunoblot by means of a patient's serum and furthermore by immunoaffinity enrichment followed by LC-MS/MS analysis. In a semi-quantitative analysis, Ara h 2 and its digestion-resistant peptides were detected in the breast milk of 9 of 32 subjects. Evidence suggests that Ara h 2 is excreted individually either rapidly (after 1, 2, 3 or 4 h) or delayed (after 8 or 12 h) and in different concentrations.Conclusions: Time and concentration of secreted Ara h 2 in breast milk appears to be individually regulated. The identification of Ara h 2 in breast milk is the prerequisite for the investigation of its sensitizing or tolerogenic properties.