Robinson M K, Horn P A, Kawabata T T, Babcock L S, Fletcher E R, Sarlo K
Procter & Gamble Company, Miami Valley Laboratories, Cincinnati, Ohio 45253, USA.
Toxicol Sci. 1998 May;43(1):39-46. doi: 10.1006/toxs.1998.2446.
A mouse intranasal test (MINT) was developed to determine the relative allergenicity of detergent enzymes. In this simple method, various doses of the enzymes are administered in a detergent matrix, via intranasal instillation, on days 1, 3, and 10, with serum samples collected on day 15. The sera are then analyzed for enzyme specific IgG1 antibody by an antigen specific enzyme immunoassay. The protease enzyme Alcalase (protease Subtilisin Carlsberg) has been used as a benchmark enzyme for development and characterization of the model. The objective of the current studies was to obtain potency comparisons with other protease and nonprotease enzymes and to begin to assess the validity of the model by comparison with potency determinations obtained with the guinea pig intratracheal (GPIT) test. The range of potencies detected among several enzymes of different classes was approximately 60-fold (compared with Alcalase). Modification of the dosing regimen to permit slightly more extended dosing did not change the relative potency determination. Comparison of data from the MINT and GPIT methods indicate that both the mouse and the guinea pig recognize the bacterial amylase Termamyl and a fungal exocellulase as more potent than Alcalase, a serine protease (Subtilisin B) and a fungal alpha-amylase (Fungamyl) as less potent than Alcalase, and the serine protease, Savinase, as equivalent to Alcalase. Also, these data are in alignment with clinical data on the prevalence of occupational enzyme sensitization. Given the simplicity and low cost of the MINT method compared with the GPIT test, these results support continued development of the method as an alternative approach for assessing the allergenicity of enzymes.
为了确定洗涤剂酶的相对致敏性，开发了一种小鼠鼻内试验（MINT）。在这个简单的方法中，在第1、3和10天通过鼻内滴注将不同剂量的酶添加到洗涤剂基质中，并在第15天采集血清样本。然后通过抗原特异性酶免疫测定法分析血清中酶特异性IgG1抗体。蛋白酶Alcalase（枯草杆菌蛋白酶卡尔伯格）已被用作该模型开发和表征的基准酶。当前研究的目的是获得与其他蛋白酶和非蛋白酶的效力比较，并通过与豚鼠气管内（GPIT）试验获得的效力测定结果进行比较，开始评估该模型的有效性。在几种不同类型的酶中检测到的效力范围约为60倍（与Alcalase相比）。调整给药方案以允许稍微延长给药时间并没有改变相对效力的测定。MINT和GPIT方法的数据比较表明，小鼠和豚鼠都认为细菌淀粉酶Termamyl和真菌外切纤维素酶比Alcalase更具效力，丝氨酸蛋白酶（枯草杆菌蛋白酶B）和真菌α-淀粉酶（Fungamyl）比Alcalase效力更低，丝氨酸蛋白酶Savinase与Alcalase效力相当。此外，这些数据与职业性酶致敏流行率的临床数据一致。鉴于与GPIT试验相比，MINT方法简单且成本低，这些结果支持继续开发该方法，作为评估酶致敏性的替代方法。
