Dearman R J, Skinner R A, Herouet C, Labay K, Debruyne E, Kimber I
Syngenta Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire SK10 4TJ, UK.
Food Chem Toxicol. 2003 Nov;41(11):1509-16. doi: 10.1016/s0278-6915(03)00167-4.
There is a growing interest in the development of methods for the evaluation of the allergenic potential of novel proteins. One approach is the measurement of specific IgE antibody production stimulated by systemic (intraperitoneal; i.p.) exposure of BALB/c strain mice. In the current investigations, inter-laboratory comparisons have been performed of IgE antibody production induced in mice by food proteins of differing sensitizing potential. Female BALB/c strain mice (n=5) were exposed to 0.1% peanut agglutinin, an allergenic constituent of peanuts, to 2% ovalbumin (OVA), a major allergenic constituent of hens' egg, or to a protein considered to lack significant allergenicity, potato agglutinin (5%). Specific IgE antibody was measured by homologous passive cutaneous anaphylaxis assay and IgG and IgG1 antibody production was analysed by enzyme-linked immunosorbent assay (ELISA). Two independent experiments were conducted in each laboratory, but with all serological analyses conducted in one of the laboratories. Each of the proteins induced vigorous IgG and IgG1 antibody responses, with no statistically significant differences in titres recorded between laboratories. Furthermore, OVA and potato agglutinin induced responses of equivalent immunogenicity with respect to both IgG and IgG1 antibody titres. Administration of peanut agglutinin and OVA each stimulated marked IgE antibody responses in every experiment. In the two laboratories, titres ranged from 1:32 and 1:64 for peanut agglutinin, and from 1:8 and 1:32 for OVA. In contrast, exposure to potato agglutinin failed to induce vigorous IgE production, with no detectable IgE (negative with neat serum), or titres of 1 (positive with neat serum only) recorded. These data demonstrate that the induction of IgE antibody by food proteins of differing allergenic potential is a relatively robust phenomenon and transferable between laboratories. Furthermore, these results provide additional evidence that the measurement of antibody (IgE) responses in BALB/c mice may allow discrimination between allergens and those materials that apparently lack allergenicity.
人们对开发评估新型蛋白质致敏潜力的方法越来越感兴趣。一种方法是测量经全身（腹腔内；i.p.）暴露BALB/c品系小鼠所刺激产生的特异性IgE抗体。在当前的研究中，已对不同致敏潜力的食物蛋白诱导小鼠产生的IgE抗体进行了实验室间比较。将雌性BALB/c品系小鼠（n = 5）暴露于0.1%的花生凝集素（花生的一种致敏成分）、2%的卵清蛋白（OVA，鸡蛋的主要致敏成分）或一种被认为缺乏显著致敏性的蛋白质——马铃薯凝集素（5%）。通过同源被动皮肤过敏试验测量特异性IgE抗体，并通过酶联免疫吸附测定（ELISA）分析IgG和IgG1抗体的产生。每个实验室进行了两项独立实验，但所有血清学分析均在其中一个实验室进行。每种蛋白质均诱导了强烈的IgG和IgG1抗体反应，各实验室记录的滴度无统计学显著差异。此外，就IgG和IgG1抗体滴度而言，OVA和马铃薯凝集素诱导的免疫原性反应相当。在每个实验中，给予花生凝集素和OVA均刺激产生了显著的IgE抗体反应。在两个实验室中，花生凝集素的滴度范围为1:32至1:64，OVA的滴度范围为1:8至1:32。相比之下，暴露于马铃薯凝集素未能诱导强烈的IgE产生，未检测到IgE（纯血清为阴性），或仅记录到滴度为1（仅纯血清为阳性）。这些数据表明，不同致敏潜力的食物蛋白诱导IgE抗体是一种相对稳定的现象，且可在实验室间转移。此外，这些结果提供了额外证据，表明在BALB/c小鼠中测量抗体（IgE）反应可能有助于区分过敏原和那些明显缺乏致敏性的物质。
