Division of Immunotoxicology, National Institute of Toxicology Research, Korea Food and Drug Administration, 122-704 Seoul, South Korea.
Environ Toxicol Pharmacol. 2004 Jun;17(2):95-102. doi: 10.1016/j.etap.2004.03.006.
The murine local lymph node assay (LLNA) has been developed as an alternative to guinea pig models for the assessment of the contact sensitization potential. However, there is a need to develop a non-radioisotopic endpoint for the LLNA, because of the radioisotopic method's requiring the use of special facilities. In this study, we investigated to evaluate the lymphocyte subpopulations in the lymph node cells following allergen and irritant treatment. Female Balb/c mice were treated by the topical application on the dorsum of both ears with sensitizers, 2,4-dinitrochlorobenzene (DNCB), toluene diisocyanate (TDI), and α-hexylcinnamaldehyde (HCA), and an irritant, sodium lauryl sulfate (SLS), once daily for three consecutive days. The lymph node (LN) cells were harvested 72h after the final treatment. Phenotypic analysis of lymphocytes subsets was performed with a flow cytometry. The allergens DNCB, TDI, and HCA and an irritant, SLS increased cell number compared to the vehicle. Mice were treated with DNCB, HCA, and TDI showed a preferential increase in the percentage of B220+CD40+ cells compared with vehicle and irritant-treated mice. There was an increase in B220+CD86+ cells of mice treated with DNCB, TDI, and HCA, but no significant increases were observed in mice treated with SLS. Mice were treated with DNCB and TDI showed an increase in the percentage of B220+CD23+ cells compared with vehicle and irritant-treated mice. These results suggest that analysis of B cell activation marker, CD40 on B cells may be useful in differentiating allergen and irritant responses in the draining lymph nodes of chemically treated mice.
小鼠局部淋巴结检测（LLNA）已被开发为评估接触致敏潜力的豚鼠模型的替代方法。然而，由于放射性同位素方法需要使用特殊设备，因此需要开发一种非放射性同位素终点的 LLNA。在这项研究中，我们研究了评估淋巴细胞亚群在淋巴结细胞中的变化，这些变化是由变应原和刺激物处理引起的。雌性 Balb/c 小鼠通过在双耳背部进行局部应用，用变应原 2,4-二硝基氯苯（DNCB）、甲苯二异氰酸酯（TDI）和α-己基肉桂醛（HCA），以及刺激性物质十二烷基硫酸钠（SLS），每日一次，连续三天进行致敏处理。在最后一次处理后 72 小时收获淋巴结（LN）细胞。使用流式细胞术对淋巴细胞亚群进行表型分析。与载体相比，变应原 DNCB、TDI 和 HCA 以及刺激性物质 SLS 增加了细胞数量。与载体和刺激性物质处理的小鼠相比，用 DNCB、HCA 和 TDI 处理的小鼠 B220+CD40+细胞的百分比增加。用 DNCB、TDI 和 HCA 处理的小鼠 B220+CD86+细胞增加，但用 SLS 处理的小鼠未观察到显著增加。与载体和刺激性物质处理的小鼠相比，用 DNCB 和 TDI 处理的小鼠 B220+CD23+细胞的百分比增加。这些结果表明，分析 B 细胞激活标志物 CD40 可能有助于区分化学处理的小鼠引流淋巴结中的变应原和刺激性反应。
