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目的探讨尘螨1类嵌合变应原TAT-Ih C-R8的致敏效果。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-Ih C-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-Ih C-R8于第0、7、14天进行腹腔注射致敏(剂量为10μg/ml);自第21天起,分别用上述致敏液(剂量为10μg/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Ig E(s Ig E)、Ig G1和Ig G2a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s Ig E和Ig G1抗体水平明显高于OVA组和R8组(P均<0.01),但其Ig G2a抗体水平则显著低于该两组(P均<0.01)。结论TAT-Ih C-R8可有效激发小鼠肺部炎症,且其致敏效果明显优于R8。
Objective To investigate the sensitization effect of dust-mite type 1 chimeric allergen TAT-Ih C-R8. Methods Forty BALB / c mice were randomly divided into three groups: PBS control group, OVA sensitization group, R8 sensitization group, and TAT-Ih C-R8 sensitized group (TIR8 group) ) 4 groups, 10 in each group. The sensitized mice were sensitized intraperitoneally with OVA, R8 and TAT-Ih C-R8 on days 0, 7 and 14 respectively (at a dose of 10 μg / ml). From day 21, Liquid (dose of 10μg / ml) inhalation atomization 30 min, continuous 7 d. The PBS group was treated with PBS for intraperitoneal and nebulized inhalation. After the last inhalation for 24 hours, the mice were sacrificed by dislocation, and bronchoalveolar lavage fluid (BLAFs) and serum were collected respectively for spleen cell culture. The levels of cytokines IFN-γ and IL-13 in BALFs and splenocyte culture supernatants (SCSs), as well as the levels of Ig E (s Ig E), Ig G1 and Ig G2a in serum were detected by ELISA BALF was counted for leukocyte and eosinophil (EOS) counts. In addition, mouse lung tissue pathological sections were made and observed after HE staining. Results Compared with the PBS group, the lung inflammation in OVA, R8 and TIR8 groups was obvious, which showed inflammatory cell infiltration, bronchial epithelial cell partial fragmentation and shedding, vasculitis and so on, while the mice in PBS group did not see Significant pathological changes. The total number of leukocytes and the number of EOS in BALF of TIR8 group were significantly higher than that of OVA and R8 groups (all P <0.01), and the levels of IL-13 in BALF and SCSs were also significantly higher than those in OVA and R8 groups ), But the content of IFN-γ was significantly lower than the two groups (all P <0.01). The serum levels of IgE and Ig G1 in TIR8 mice were significantly higher than those in OVA and R8 groups (all P <0.01), but the levels of Ig G2a antibody in TIR8 mice were significantly lower than those in both groups (P <0.01). Conclusion TAT-Ih C-R8 can effectively stimulate lung inflammation in mice, and its sensitization effect is significantly better than R8.