Chin J Schisto Control ›› 2015, Vol. 27 ›› Issue (5): 485-.DOI: 10.16250/j.32.1374.2015071

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Analysis of sensitization effect of chimeric allergen TAT-IhC-R8 derived from major allergen group 1 genes of dust mites

LI Chao-pin* | ZHAO Bei-bei|JIANG Yu-xin|DIAO Ji-dong|LI Na| LU Wei   

  1. Department of Medical Parasitology|Wannan Medical College| Wuhu 241002| China
  • Online:2015-10-21 Published:2015-10-21

尘螨1类嵌合变应原TAT?IhC?R8的致敏效果分析

李朝品*|赵蓓蓓|姜玉新|刁吉东|李娜|陆维   

  1. 皖南医学院医学寄生虫学教研室 (芜湖 241002)
  • 作者简介:李朝品| 男| 博士| 教授。研究方向: 病原生物学与免疫学
  • 基金资助:
    国家自然科学基金 (81270091、 30872367、 81172790)

Abstract: Objective Objective To explore the sensitization effect of allergen TAT? IhC?R8,derived from major allergen group 1 genes of dust mites. Methods Methods Forty BALB/c mice were randomly divided into 4 groups,namely PBS group,ovalbumin (OVA)group,R8 group and TAT?IhC?R8(TIR8)group,10 mice each group. All the mice in OVA,R8 and TIR8 groups were treated with corresponding allergens(10 μg/ml)on the 0,7th and 14th day by intraperitoneal injection and nebulized inhalation on day 21 with the concentration of 30 min/d for 7 days. The mice in PBS group were treated with PBS. Twenty?four hours after the last challenge,all the mice were sacrificed,their bronchoalveolar lavage fluids(BALFs)and sera were collected and their spleen cells were cultured. ELISA was performed to detect the levels of IFN?γ and IL?13 in BALFs and supernatants of cultured splenocytes(SCSs)of the mice,as well as the levels of allergen?specific IgE(sIgE),IgG1 and IgG2a in their sera. The number of white blood cells and eosinophils in BALF were calculated. In addition,the airway inflammation and mucus secretion were ana? lyzed by haematoxylin and eosin(H&E)staining. Results Results Compared with the PBS group,the lung inflammations of mice in the OVA,R8 and TIR8 groups were observed obviously,including inflammatory infiltration,bronchial epithelial cell breakage and falling off,as well as vasculitis. The numbers of the total white blood cells and eosinophils in BALF of mice in the TIR8 group were significantly more than those in the OVA and R8 groups(all P < 0.01) . The IL?13 levels in BALFs and SCSs of mice in the TIR8 group were significantly higher than those in the OVA group and R8 group(all P < 0.01) . However,the level of IFN? γ of mice in the TIR8 group was lower than those in the latter 2 groups(all P < 0.01) . In addition,the levels of sera sIgE and IgG1 of mice from the TIR8 group were significantly higher than those in the OVA group and R8 group(all P < 0.01),but the level of IgG2a of the former was significantly lower than those of the latter groups(all P < 0.01) . Conclusions Conclusions TAT?IhC?R8 can effectively stimulate lung inflammations of mice, and its sensitization effect is better than R8’ s.

Key words: Dust mite;Chimeric allergen;Allergic asthma;Animal model;TAT?IhC?R8;R8

摘要: 目的 目的 探讨尘螨1类嵌合变应原TAT?IhC?R8的致敏效果。方法 方法 将40只BALB/c小鼠随机分为PBS对照组 (PBS组)、 OVA致敏组 (OVA组)、 R8致敏组 (R8组) 和TAT?IhC?R8致敏组 (TIR8组) 4组, 每组10只。上述3个致敏组分 别用OVA、 R8和TAT?IhC?R8于第0、 7、 14 天进行腹腔注射致敏 (剂量为10 μg/ml); 自第21天起, 分别用上述致敏液 (剂量 为10 μg/ml) 雾化吸入30 min, 连续7 d。PBS组则用PBS进行腹腔注射和雾化吸入。最后一次雾化吸入24 h后将小鼠脱 臼处死, 分别收集支气管肺泡灌洗液 (BLAFs) 和血清, 并进行脾细胞培养。以ELISA法检测BALFs及脾细胞培养上清液 (SCSs) 中细胞因子IFN?γ和IL?13水平, 以及血清中特异性抗体IgE (sIgE)、 IgG1和IgG2a水平; 并对BALF进行白细胞和嗜 酸性粒细胞 (EOS) 计数。此外, 制作小鼠肺组织病理切片, 经HE染色后进行观察。结果 结果 与PBS组相比, OVA、 R8组和 TIR8组小鼠肺部炎症均明显, 表现为肺组织炎性细胞浸润、 支气管上皮细胞部分断裂及脱落、 血管炎等, 而PBS组小鼠未 见明显病理改变。TIR8组小鼠BALF中白细胞总数和EOS数均显著高于OVA组和R8组 (P 均< 0.01), 其BALF及SCSs 中IL?13水平亦明显高于OVA组和R8组 (P 均< 0.01), 但IFN?γ含量则显著低于该两组 (P 均< 0.01)。TIR8组小鼠血清 sIgE和IgG1抗体水平明显高于OVA组和R8组 (P 均< 0.01), 但其IgG2a抗体水平则显著低于该两组 (P 均< 0.01)。结论 结论 TAT?IhC?R8可有效激发小鼠肺部炎症, 且其致敏效果明显优于R8。

关键词: 尘螨; 嵌合变应原; 过敏性哮喘; 动物模型; TAT?IhC?R8; R8

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