Tarkowski Maciej, Kur Barbara, Polakowska Ewa, Jabłońska Ewa
Department of Immunotoxicology, Nofer Institute of Occupational Medicine, Lódź, Poland.
Int J Occup Med Environ Health. 2008;21(3):253-62. doi: 10.2478/v10001-008-0031-y.
To investigate immunological changes in lymph nodes based on expression of cell-specific receptors and cytokine expression profile and accompanying inflammatory reactions in lungs of mice treated with chemicals of known potentials to induce respiratory sensitization and those in which activity in this regard is unclear.
On day 1 and 7, Balb/c mice received toluene-2,4-diisocyanate (TDI), trimellitic anhydride (TMA), 1-chloro-2,4-dinitrobenzene (DNCB), glutaraldehyde (GA), formaldehyde (FA), benzalkonium chloride (ChB) or vehicle. On day 14, they received a single intranasal instillation with the same chemical or vehicle. On day 15, auricular lymph nodes (LN) were excised and used for analyzes of T-, B-cells, expression of CD44 and for the estimation of IL-4 and IFN-gamma production after in vitro stimulation with concanavalin A (ConA) and also for IL-4 and IFN-gamma mRNA expression analyses using Real-Time PCR. Inflammatory changes in lungs were observed by estimation of TNF-alpha and MIP-2 concentrations and cell numbers and their type in BAL.
There were no significant changes in cell subpopulations of T helper cells in LN. The percent of B cells was significantly increased after treatment with DNCB, TDI, and GA. Increased expression of CD44 on T cells was also observed. Both IL-4 and IFN-gamma were found increased in TDI- and FA-treated mice, while only IL-4 was increased in TMA-treated mice. Real-Time PCR analyses, however, showed increased IL-4 mRNA expression for TDI- and TMA-, and IFN-gamma mRNA expression for DNCB-treated mice. We haven't observed significant changes in inflammatory reactions in the lungs of exposed animals.
Studying immunological changes with first determining the activation status of T cells followed by analyzes of expression of mRNA for Th1 and Th2 cytokines in murine model could be a useful method for assessment of the potentials of chemicals to induce respiratory sensitization but is not sufficient. Addition of ventilatory measurements, but not necessarily inflammatory reactions, could complete the model.
基于细胞特异性受体的表达、细胞因子表达谱以及伴有炎症反应，研究已知具有诱导呼吸道致敏潜力的化学物质处理的小鼠以及在这方面活性不明确的化学物质处理的小鼠的淋巴结免疫变化及肺部情况。
在第1天和第7天，Balb/c小鼠接受甲苯-2,4-二异氰酸酯（TDI）、偏苯三酸酐（TMA）、1-氯-2,4-二硝基苯（DNCB）、戊二醛（GA）、甲醛（FA）、苯扎氯铵（ChB）或赋形剂处理。在第14天，它们接受相同化学物质或赋形剂的单次鼻内滴注。在第15天，切除耳淋巴结（LN），用于分析T细胞、B细胞、CD44的表达，并用于评估在体外经伴刀豆球蛋白A（ConA）刺激后IL-4和IFN-γ的产生，以及使用实时PCR分析IL-4和IFN-γ mRNA的表达。通过估计TNF-α和MIP-2浓度以及支气管肺泡灌洗（BAL）中的细胞数量及其类型来观察肺部的炎症变化。
LN中辅助性T细胞的细胞亚群没有显著变化。用DNCB、TDI和GA处理后，B细胞百分比显著增加。还观察到T细胞上CD44表达增加。在TDI和FA处理的小鼠中，IL-4和IFN-γ均升高，而在TMA处理的小鼠中只有IL-4升高。然而，实时PCR分析显示，TDI和TMA处理的小鼠IL-4 mRNA表达增加，DNCB处理的小鼠IFN-γ mRNA表达增加。我们没有观察到暴露动物肺部炎症反应的显著变化。
在小鼠模型中，先确定T细胞的激活状态，然后分析Th1和Th2细胞因子mRNA的表达来研究免疫变化，可能是评估化学物质诱导呼吸道致敏潜力的一种有用方法，但并不充分。增加通气测量，而不一定是炎症反应，可以完善该模型。
