Lee Jong Kwon, Park Jae Hyun, Eom Juno H, Kim Hyung Soo, Oh Hye Young
Division of Immunotoxicology, National Institute of Toxicology Research, Korea Food and Drug Administration, 122-704 Seoul, South Korea.
Environ Toxicol Pharmacol. 2005 Jul;20(1):225-32. doi: 10.1016/j.etap.2005.02.002. Epub 2005 Mar 23.
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 populations of intracellular cytokine producing cells and to analyze the expression of mRNA levels in the lymph node (LN) cells following allergen and irritant. Female Balb/c mice were treated by the topical application on the dorsum of both ears with strong sensitizers, 2,4-dinitrochlorobenzene (DNCB) and toluene diisocyanate (TDI) and a strong irritant, sodium lauryl sulfate (SLS), once daily for 3 consecutive days. The lymph node cells were harvested 72h after the final treatment. The analysis of intracellular cytokine cell in LN cells was performed with a flow cytometry. Mice were treated with DNCB and TDI showed a preferential increase in the percentage of CD4+IL-2+ cells compared with vehicle and irritant-treated mice. There was an increase in CD4+IFN-g+ cells of mice treated with DNCB and TDI, 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 CD4+IL-4+ cells compared with vehicle and irritant-treated mice. There was an increase in the mRNA level for interleukin 4 (IL-4) in mice treated with DNCB and TDI, but no significant increases were observed in mice treated with SLS. These results suggest that the population of interferon-gamma (IFN-g+) and IL-4+ cells on CD4+ cells and the mRNA expression for IL-4 in lymphocytes could be selectively modulated in allergen-treated mice.
小鼠局部淋巴结试验（LLNA）已被开发出来，作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施，因此有必要为LLNA开发一种非放射性同位素终点。在本研究中，我们调查了评估细胞内细胞因子产生细胞群体，并分析变应原和刺激物作用后淋巴结（LN）细胞中mRNA水平的表达。雌性Balb/c小鼠通过在双耳背部局部涂抹强致敏剂2,4-二硝基氯苯（DNCB）和甲苯二异氰酸酯（TDI）以及强刺激剂十二烷基硫酸钠（SLS）进行处理，连续3天每天一次。在最后一次处理后72小时收集淋巴结细胞。用流式细胞术对LN细胞中的细胞内细胞因子进行分析。与赋形剂和刺激物处理的小鼠相比，用DNCB和TDI处理的小鼠CD4+IL-2+细胞百分比优先增加。用DNCB和TDI处理的小鼠CD4+IFN-γ+细胞增加，但用SLS处理的小鼠未观察到显著增加。与赋形剂和刺激物处理的小鼠相比，用DNCB和TDI处理的小鼠CD4+IL-4+细胞百分比增加。用DNCB和TDI处理的小鼠白细胞介素4（IL-4）的mRNA水平增加，但用SLS处理的小鼠未观察到显著增加。这些结果表明变应原处理的小鼠中CD4+细胞上的干扰素-γ（IFN-γ+）和IL-4+细胞群体以及淋巴细胞中IL-4的mRNA表达可被选择性调节。
