Department of Toxicology, National Institute of Toxicological Research, Korea Food and Drug Administration, Seoul 122-704, Republic of Korea.
Environ Toxicol Pharmacol. 2003 Jun;14(1-2):61-8. doi: 10.1016/S1382-6689(03)00025-5.
The mouse ear swelling test (MEST) was developed as an alternative to guinea pig models for measuring the contact sensitization potential. However, the MEST relies on the quantitative measurement of ear swelling by micrometer as the means of determining the endpoint. The purpose of this study was to investigate the possibility of using cell proliferation in the ear and lymph node by bromodeoxyuridine (BrdU) immunohistochemistry as a reliable marker for MEST. Female Balb/c mice were treated by the topical application of various sensitizers, 2,4-dinitrochlorobenzene (DNCB), toluene diisocyanate (TDI) and α-hexylcinnamaldehyde (HCA) and an irritant, sodium lauryl sulfate (SLS) following the protocol of MEST. The proliferation of cells in the ear and auricular lymph node was analyzed by BrdU incorporations into cells. There were significant increases in the cell proliferations of the ear and auricular lymph node in mice treated with DNCB and TDI compared to the vehicle control. All allergens and the irritant were correctly identified by the MEST using BrdU immunohistochemistry of lymph node responses. The standard MEST assay showed positive results in the case of the strong sensitizers, DNCB and TDI. However, HCA and SLS were not correctly identified in the ear swelling assay. These results suggest that the measurement of cell proliferation in the auricular lymph node using BrdU immunohistochemistry could provide a reliable marker for MEST.
小鼠耳肿胀试验（MEST）是作为豚鼠模型的替代方法开发的，用于测量接触致敏潜能。然而，MEST 依赖于通过测微计定量测量耳肿胀作为确定终点的手段。本研究的目的是研究使用耳和淋巴结中的细胞增殖通过溴脱氧尿苷（BrdU）免疫组织化学作为 MEST 的可靠标志物的可能性。雌性 Balb/c 小鼠通过用各种敏化剂（2,4-二硝基氯苯（DNCB），甲苯二异氰酸酯（TDI）和α-己基肉桂醛（HCA））和刺激性物质（十二烷基硫酸钠（SLS））处理，根据 MEST 的方案进行局部应用。通过 BrdU 掺入细胞来分析耳和耳淋巴结中的细胞增殖。与载体对照相比，用 DNCB 和 TDI 处理的小鼠的耳和耳淋巴结中的细胞增殖显着增加。使用淋巴结反应的 BrdU 免疫组织化学，所有变应原和刺激性物质均被 MEST 正确识别。标准 MEST 测定法对强敏化剂 DNCB 和 TDI 显示阳性结果。然而，在耳肿胀测定中，HCA 和 SLS 未被正确识别。这些结果表明，使用 BrdU 免疫组织化学测量耳淋巴结中的细胞增殖可以为 MEST 提供可靠的标志物。
