Development of a peptide reactivity assay for screening contact allergens
Development of a peptide reactivity assay for screening contact allergens
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DOI：
10.1093/toxsci/kfh213
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发表时间：
2004-10-01
影响因子：
3.8
通讯作者：
Lepoittevin, JP
中科院分区：
文献类型：
--
作者：
Gerberick, GF;Vassallo, JD;Lepoittevin, JP
皮肤致敏引起的过敏性接触性皮炎是一种常见的职业和环境健康问题。近年来，局部淋巴结检(LLNA)已成为评估化学物质皮肤致敏潜力的实用方法。除了准确识别皮肤致敏剂外，LLNA还可以提供相对致敏效力的可靠测量；这些信息对于成功管理人类健康风险至关重要。然而，即使LLNA提供了显著的动物福利好处，人们仍然对开发用于皮肤致敏测试的非动物测试方法感兴趣。化学过敏原的一个特征是在诱导皮肤过敏之前与蛋白质发生反应的能力。大多数化学过敏原是亲电性的，因此与半胱氨酸或赖氨酸等亲核氨基酸发生反应。为了确定反应性是否与致敏潜力相关，对38种代表不同效力(从弱到极端)的过敏原和非致敏剂的化学物质与谷胱甘肽或三种含有半胱氨酸、赖氨酸或组氨酸的合成肽的反应能力进行了评估。在谷胱甘肽的反应时间为15分钟或三个合成肽的反应时间为24小时后，用高效液相色谱法对样品进行分析。UV检测用于监测谷胱甘肽或反应后的多肽的耗竭。结果表明，变应原效力与谷胱甘肽(p=0.001)、赖氨酸(p=0.025)和半胱氨酸(p=0.020)的耗竭之间存在显著的相关性(Spearman相关)，但与组氨酸的耗竭无关。半胱氨酸敏感性最高(80.8%)，组氨酸敏感性最低(11.5%)。提供的数据表明，测量多肽的反应性对于筛选化学物质的皮肤致敏效力具有实用价值，从而有可能减少我们对动物试验方法的依赖。
Allergic contact dermatitis resulting from skin sensitization is a common occupational and environmental health problem. In recent years, the local lymph node assay (LLNA) has emerged as a practical option for assessing the skin sensitization potential of chemicals. In addition to accurate identification of skin sensitizers, the LLNA can also provide a reliable measure of relative sensitization potency; information that is pivotal in successful management of human health risks. However, even with the significant animal welfare benefits provided by the LLNA, there is still interest in the development of nonanimal test methods for skin sensitization testing. One characteristic of a chemical allergen is its ability to react with proteins prior to the induction of skin sensitization. The majority of chemical allergens is electrophilic and as such reacts with nucleophilic amino acids like cysteine or lysine. In order to determine if reactivity correlates with sensitization potential, 38 chemicals representing allergens of different potencies (weak to extreme) and nonsensitizers were evaluated for their ability to react with glutathione or three synthetic peptides containing either cysteine, lysine, or histidine. Following a 15-min reaction time for glutathione or a 24 h reaction period for the three synthetic peptides, the samples were analyzed by HPLC. UV detection was used to monitor the depletion of glutathione or the peptide following reaction. The results demonstrate that a significant correlation (Spearman correlation) exists between allergen potency and the depletion of glutathione (p = 0.001), lysine (p = 0.025), and cysteine (p = 0.020), but not histidine. The peptide with the highest sensitivity was cysteine (80.8%) whereas histidine was the least sensitive (11.5%). The data presented show that measuring peptide reactivity has utility for screening chemicals for their skin sensitization potency and thus potential for reducing our reliance on animal test methods.