Study on the identification of industrial respiratory allergens using the analysis of immunity in lung.

本研究旨在探讨职业性呼吸道变应原致敏肺的免疫功能分析是否能为职业性呼吸道变应原致敏肺提供正确的评价。首先，在小鼠剃光的背部皮肤上涂上甲苯二异氰酸酯（TDI），这是一种有效的呼吸敏感性。末次染毒后2周，小鼠鼻内接种TDI，分别于染毒后0、1、6、12 h，采用RT-PCR方法检测肺组织细胞因子基因表达，结果显示，未致敏和致敏小鼠肺组织均检测到IL-2、IL-5和IFN-γ mRNA，而致敏和激发小鼠肺组织仅检测到IL-4 mRNA。这些发现也观察到在吸入TDI致敏和激发的小鼠中。此外，我们已经检查了四种细胞因子mRNA的基因表达模式是否可以区分呼吸道过敏原和接触性过敏原。小鼠被涂上苦基氯（PCI），这是一种有效的接触敏感性。在最后一次暴露后2周，小鼠鼻内接种PCI进行激发，结果发现，在激发后6 h，PCI致敏小鼠肺组织中IL-4和IFN-γ mRNA的基因表达较TDI致敏小鼠弱，因此，RT-PCR分析细胞因子谱可用于评价致敏物的致敏性。

In this study, we have examined whether the analysis of immune function in lung sensitized with occupational respiratory allergens could provide the proper evaluation for them. Firstly, mice were painted on the shaved back skin with toluene diisocyanate (TDI), known as a potent respiratory sensitivity. Two weeks after the last exposure, mice were challenged with intranasal inoculation of TDI.After 0,1,6 and 12 hours, the gene expression of cytokine was analyzed in the lung of mice using RT-PCR.It was observed that IL-2, IL-5 and IFN-gamma mRNAs were detected in non-sensitized and sensitized lung of mice, but IL-4 mRNA was detected only in sensitized and challenged lung of mice. These finding were also observed in mice sensitized and challenged with inhaled TDI.Moreover, we have examined whether the pattern of the gene expression of four cytokine mRNAs could distinguish respiratory allergens from contact allergens. Mice were painted with picryl chloride (PCI), known as a potent contact sensitivity. Two weeks after the last exposure, mice were challenged with intranasal inoculation of PCI.It was observed that the gene expression of IL-4 and IFN-gamma mRNAs were extremely weak in the lungs of mice sensitized with PCI compared with those of TDI 6 hours after challenging.Therefore, the analysis of cytokine profiles using RT-PCR was suggested to be available for the evaluation of sensitizers causing respiratory allergy.