Fujiwara Shunsuke, Yasunaga Sho, Iwabuchi Shigehiro, Masuda Kenichi, Ohno Koichi, Tsujimoto Hajime
Department of Veterinary Internal Medicine, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
Vet Immunol Immunopathol. 2003 May 30;93(1-2):9-20. doi: 10.1016/s0165-2427(03)00049-7.
Japanese cedar (Cryptomeria japonica, CJ) pollinosis is mediated by type-I hypersensitivity and induces seasonal rhinitis and conjunctivitis in humans. Previous studies showed that dogs could be experimentally sensitized with CJ pollen. In this study, we carried out quantitative analysis of mRNA levels of various cytokines in the peripheral blood mononuclear cells (PBMC) of 12 dogs experimentally sensitized to Japanese cedar pollen. Experimental sensitization was carried out by injection of crude CJ pollen extract with aluminium hydroxide gel. The expression levels of interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, IL-18, interferon (IFN)-gamma, transforming growth factor (TGF)-beta(1), and tumor necrosis factor (TNF)-alpha mRNAs in the PBMC were quantified using a real-time sequence detection system. In the PBMC tested without culture, the expression levels of IL-8 and TNF-alpha mRNAs in experimentally sensitized dogs were significantly higher than those in control dogs. The expression level of IFN-gamma mRNA in the sensitized group was significantly lower than that in the control group. When the PBMCs were cultured in the presence of CJ pollen extract, the level of IL-4 mRNA expression was markedly increased in the PBMC from the experimentally sensitized dogs. In the PBMC stimulated with the CJ pollen extract, the expression level of IL-2 mRNA in the sensitized group was also significantly higher than that in the control group. Our data indicated that a Th2 response and proliferation of PBMC occur in response to the sensitizing antigen in dogs experimentally sensitized with CJ pollen, and revealed the presence of antigen-specific Th2 cells in this canine model. In addition, the expression levels of the mRNAs encoding proinflammatory cytokines were shown to be elevated after CJ pollen sensitization, indicating the activation of monocytes and macrophages.
日本柳杉(Cryptomeria japonica,CJ)花粉症由I型超敏反应介导,可诱发人类季节性鼻炎和结膜炎。既往研究表明,犬可通过实验性致敏于CJ花粉。在本研究中,我们对12只经实验性致敏于日本柳杉花粉的犬外周血单个核细胞(PBMC)中多种细胞因子的mRNA水平进行了定量分析。通过注射含氢氧化铝凝胶的CJ花粉粗提物进行实验性致敏。使用实时序列检测系统对PBMC中白细胞介素(IL)-1β、IL-2、IL-4、IL-6、IL-8、IL-10、IL-18、干扰素(IFN)-γ、转化生长因子(TGF)-β1和肿瘤坏死因子(TNF)-α mRNA的表达水平进行定量。在未经培养的PBMC检测中,实验性致敏犬中IL-8和TNF-α mRNA的表达水平显著高于对照犬。致敏组中IFN-γ mRNA的表达水平显著低于对照组。当PBMC在CJ花粉提取物存在的情况下培养时,实验性致敏犬的PBMC中IL-4 mRNA的表达水平显著增加。在用CJ花粉提取物刺激的PBMC中,致敏组中IL-2 mRNA的表达水平也显著高于对照组。我们的数据表明,在用CJ花粉实验性致敏的犬中,PBMC会针对致敏抗原发生Th2反应和增殖,并揭示了该犬模型中存在抗原特异性Th2细胞。此外,编码促炎细胞因子的mRNA表达水平在CJ花粉致敏后升高,表明单核细胞和巨噬细胞被激活。