School of Biological Sciences, University of Nebraska, Lincoln, NE, USA; Food Allergy Research and Resource Program, Dept. of Food Science and Technology, University of Nebraska, Lincoln, NE, USA; The Translational Pulmonary and Immunology Research Center (TPIRC), Long Beach, CA, USA.
Food Allergy Research and Resource Program, Dept. of Food Science and Technology, University of Nebraska, Lincoln, NE, USA.
Food Chem Toxicol. 2020 Jul;141:111398. doi: 10.1016/j.fct.2020.111398. Epub 2020 May 11.
Currently no validated animal model is predictive of human responses in ranking purified dietary proteins in the prevalence or potency of food allergy in humans. Since the gastrointestinal microbiota is thought to influence oral tolerance, we hypothesize that a germ-free mouse model will more accurately predict atopic human responses than conventional mice. Germ-free C3H/HeN mice were immunized with 60 μg Ara h 2, BLG, or LOX by three weekly intraperitoneal (IP) injections with alum adjuvant. One week following the final immunization an IP challenge of 500 μg of Ara h 2, BLG, or LOX was administered. Thirty minutes post-challenge clinical scores were graded and body temperatures recorded. The presence of protein-specific IgE and mast cell protease concentrations in mouse sera were determined using ELISA. Upon challenge germ-free mice sensitized with Ara h 2 and BLG exhibited significantly more severe clinical scores compared to germ-free mice immunized with LOX. Hypothermic responses in challenged mice differed between the three proteins post-challenge. Results indicate that this model can differentiate between potent and non-allergens based on temperature drop, clinical scores, and biomarkers. Additional proteins with known human exposure and allergenicity are needed to confirm the predictive accuracy.
目前没有经过验证的动物模型可以预测人类对纯化膳食蛋白质在人类食物过敏的普遍性或强度的反应。由于胃肠道微生物群被认为会影响口服耐受，我们假设无菌小鼠模型将比常规小鼠更能准确预测特应性人类的反应。无菌 C3H/HeN 小鼠通过每周三次腹腔内（IP）注射含明矾佐剂的 60μg Ara h 2、BLG 或 LOX 进行免疫。在最后一次免疫后一周，进行 500μg Ara h 2、BLG 或 LOX 的 IP 挑战。挑战后 30 分钟进行临床评分，并记录体温。使用 ELISA 测定小鼠血清中蛋白质特异性 IgE 和肥大细胞蛋白酶浓度。在挑战中，与用 LOX 免疫的无菌小鼠相比，用 Ara h 2 和 BLG 敏化的无菌小鼠表现出更严重的临床评分。挑战后三种蛋白质的小鼠体温反应不同。结果表明，该模型可以根据体温下降、临床评分和生物标志物区分有效原和非过敏原。需要具有已知人类暴露和致敏性的其他蛋白质来确认预测的准确性。
