INTRODUCTION This 4-center study is part of a project to validate a food allergy murine model for safety testing of hydrolyzed infant formulas. AIM The aim of the current multi-center experiment was to evaluate the residual allergenicity of three partial hydrolyzed whey proteins (pWH) in a multiple-parameter cow's milk allergy murine model and to compare to the classically used guinea pig model. Previous work showed differences in the magnitude of the allergic response to whey between centers. To get a first insight in the effect of housing on the robustness of the mouse model, microbiota composition of non-sensitized mice was analyzed and compared between centers. METHODS Mice were sensitized intragastrically (i.g.) with whey, pWH or eWH using cholera toxin as an adjuvant. In mice, whey-IgE/IgG1, acute allergic symptoms were determined upon whey challenge. Guinea pigs were orally sensitized ad libitum via the drinking water (day 0-37) and challenged intravenously with whey on day 49. The microbial composition in fecal samples was determined in non-sensitized mice in all 4 research centers before and after conduct of the study. RESULTS Elevated levels of whey-IgG1 were detected in whey-sensitized mice in all centers. Except for pWH-A in center 4, we observed elevated levels of whey-IgE in whey-sensitized mice and mice sensitized with pWH-A, -B, -C. Center 2 was excluded from further analysis because of non-significant IgE levels in the positive control. In contrast to whey-mice, pWH-A treated mice showed no acute skin response, mMCP-1 release or change in body temperature upon whey challenge in all centers, which corresponds with the absence of anaphylactic shock symptoms in both the mouse and guinea pig model. pWH-B and -C induced anaphylactic shock symptoms in the guinea-pig and mice whereas results on the remaining allergic outcomes in mice were inconclusive. No differences in microbiota composition were measured in response to the challenge and Microbiota composition depended on the location of the centers. CONCLUSIONS Both animal models showed comparable results on the residual allergenicity of partial hydrolyzed whey proteins, but none of the centers was able to differentiate between the residual sensitizing capacities of the pWH-B and -C based on a single elicitation parameter in the murine model. Differences in microbiota composition might contribute to the robustness of the food allergy murine model. For a well-balanced prediction on the potential allergenicity of hydrolyzed infant formulas a multiple murine parameter model is suggested to decrease the risk of false positive or false negative results. A future challenge is to develop an overall scoring system for proper risk assessment, taking all parameters into account.

中文翻译：

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多中心评估比较部分水解乳清蛋白在牛奶过敏小鼠模型中的残留过敏性——与单参数豚鼠模型的比较

简介 这项 4 中心研究是验证用于水解婴儿配方食品安全性测试的食物过敏鼠模型项目的一部分。目的 当前多中心实验的目的是评估多参数牛奶过敏鼠模型中三种部分水解乳清蛋白 (pWH) 的残留过敏性，并与经典使用的豚鼠模型进行比较。以前的工作表明，不同中心对乳清的过敏反应程度存在差异。为了初步了解住房对小鼠模型稳健性的影响，我们对非致敏小鼠的微生物群组成进行了分析，并在中心之间进行了比较。方法 使用霍乱毒素作为佐剂，用乳清、pWH 或 eWH 对小鼠进行胃内 (ig) 致敏。在小鼠中，乳清-IgE/IgG1、根据乳清挑战确定急性过敏症状。豚鼠通过饮用水（第 0-37 天）口服致敏，并在第 49 天用乳清进行静脉注射。在所有 4 个研究中心的未致敏小鼠中测定粪便样本中的微生物组成。学习。结果 在所有中心的乳清致敏小鼠中均检测到乳清-IgG1 水平升高。除了中心 4 中的 pWH-A，我们观察到乳清致敏小鼠和 pWH-A、-B、-C 致敏小鼠的乳清 IgE 水平升高。由于阳性对照中 IgE 水平不显着，中心 2 被排除在进一步分析之外。与乳清小鼠相比，pWH-A 处理的小鼠在所有中心均未表现出急性皮肤反应、mMCP-1 释放或体温变化。这与小鼠和豚鼠模型中均不存在过敏性休克症状相对应。pWH-B 和 -C 在豚鼠和小鼠中诱发过敏性休克症状，而对小鼠其余过敏结果的结果尚无定论。没有测量到响应挑战时微生物群组成的差异，微生物群组成取决于中心的位置。结论 两种动物模型在部分水解乳清蛋白的残留过敏性方面都显示出类似的结果，但没有一个中心能够根据小鼠模型中的单个诱发参数区分 pWH-B 和 -C 的残留致敏能力。微生物群组成的差异可能有助于食物过敏小鼠模型的稳健性。为了对水解婴儿配方奶粉的潜在过敏性进行均衡预测，建议使用多鼠参数模型来降低假阳性或假阴性结果的风险。未来的挑战是为适当的风险评估开发一个整体评分系统，同时考虑所有参数。