Development of non-radio isotopic endpoint of murine local lymph node assay based on 5-bromo-2′-deoxyuridine (BrdU) incorporation
Development of non-radio isotopic endpoint of murine local lymph node assay based on 5-bromo-2′-deoxyuridine (BrdU) incorporation
复制标题
DOI：
10.1016/s0378-4274(00)00315-5
复制
发表时间：
2001-03-08
影响因子：
3.5
通讯作者：
Kimber, I
中科院分区：
文献类型：
--
作者：
Takeyoshi, M;Yamasaki, K;Kimber, I
过敏性接触性皮炎是一个严重的健康问题。在过去的十年中，已经开发出鼠局部淋巴结测定法（LLNA）来检测化学过敏原，并进行了国际验证研究。我们试图通过使用5-bromo-2'-脱氧尿苷（BRDU）掺入来建立LLNA的替代性非放射性终点，以代替标准方法中采用的[H-3]胸苷等放射性异位素。 BRDU每天连续三天以5 mg/动物为单一给药。测试化学物质的应用。使用酶免疫吸附测定技术测量BRDU融合到排水淋巴结细胞中。在这项研究中，将P-苯甲喹酮（PBQ），三核酸酸酐（TMA），Citral（CT）和Dextran（Dex）用作试验性化学物质。 PBQ，TMA和CT在豚鼠最大化测试中被归类为中度至强敏机，并且在原始LLNA中呈阳性，也发现使用BRDU掺入在替代LLNA中引起正反应。相反，DEX在与先前的豚鼠和LLNA数据一致的修饰测定中测试了阴性。因此，在需要避免使用放射性同位素的情况下，使用BRDU掺入的修改的LLNA端点可能代表了标准测定法的有用替代方案。 （c）2001 Elsevier Science Ireland Ltd.保留所有权利。
Allergic contact dermatitis is a serious health problem. Over the last decade, the murine local lymph node assay (LLNA) has been developed to detect chemical allergens, and international validation studies have been conducted. We have tried to establish an alternative non-radioisotopic endpoint for the LLNA by using 5-bromo-2'-deoxyuridine (BrdU) incorporation in place of radioisotopes, such as [H-3]thymidine, employed in the standard method. BrdU was given as a single administration at 5 mg/animal 2 days following three consecutive daily. applications of a test chemical. BrdU incorporation into draining lymph node cells was measured using an enzyme immunosorbent assay technique. In this study, p -benzoquinone(PBQ), trimellitic anhydride (TMA), citral(CT) and dextran (DEX) were used as pilot chemicals. PBQ, TMA and CT, which are classified as moderate to strong sensitizers in the guinea pig maximization test and were positive in the original LLNA, were also found to elicit positive responses in the alternative LLNA using BrdU incorporation. In contrast, DEX tested negative in the modified assay consistent with previous guinea pig and LLNA data. Consequently, the modified LLNA endpoint using BrdU incorporation may represent a useful alternative to the standard assay in situations, where there is a need to avoid the use of radioisotopes. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.