小鼠局部变应性鼻炎耐受模型的研究

Establishment of local allergic rhinitis tolerance in mouse model

目的:通过鼻腔滴入过敏原建立小鼠局部变应性鼻炎动物耐受模型,并对其免疫学指标及特征进行研究。方法:使用卵清蛋白(OVA)和磷酸盐缓冲液(PBS)对小鼠每日局部滴鼻,记录过敏症状,检测小鼠血清OVA特异性抗体(IgE、IgG1、IgG2a)及脾脏细胞培养上清的细胞因子(IL-4、IL-10、IFN-γ)浓度,观察鼻黏膜嗜酸粒细胞及杯状细胞的浸润情况,用RNA-seq技术分析局部鼻黏膜基因的变化。结果:小鼠在OVA刺激下首先表现为过敏症状加重,血清OVA特异性抗体IgE、IgG1、IgG2a和体外培养脾细胞上清中IL-4、IFN-γ、IL-10升高,鼻黏膜嗜酸粒细胞和杯状细胞显著升高,鼻黏膜编码IL-10、TGF-β基因表达上调,嗜酸粒细胞活化基因上调。随着滴鼻时间继续延长,过敏症状逐渐减轻,血清OVA特异性抗体IgE、IgG1、IgG2a则持续增加,体外培养脾脏细胞上清IL-4下降,IL-10升高,IFN-γ有下降的趋势,鼻黏膜杯状细胞显著减少,嗜酸粒细胞活化基因显著下调,鼻黏膜编码IL-10、TGF-β等耐受基因维持高表达。筛选出10个与局部变应性鼻炎免疫耐受相关的核心基因:Rps27a、Uba52、Kng2、Gnal、C3、Rtp4、Reep1、Rtp2、Rtp1、Reep5。结论:通过局部持续滴入过敏原,可使小鼠先形成过敏,继而产生免疫耐受。这种免疫耐受的产生,可能是在变应原的持续作用下,诱导了全身及局部的免疫耐受效应。

Objective: To establish a mouse model of local allergic rhinitis tolerance by intranasal infusion of allergen, and study its immunological indexes and characteristics. Methods: The mice were given ovalbumin(OVA) and phosphate buffer solution(PBS) daily, and their allergic symptoms were recorded. OVA-specific antibodies(IgE, IgG1, IgG2 a) in serum and cytokine(IL-4, IL-10, IFN-γ) in splenic culture supernatant were detected. The infiltration of eosinophils and goblet cells in nasal mucosa were observed, and the changes in local nasal mucosa genes were analyzed by RNA-seq technique. Results: Mice first showed aggravation of allergic symptoms when stimulated with OVA. The serum OVA-specific antibodies IgE, IgG1, IgG2 a and the cytokine(IL-4, IL-10, IFN-γ) Iin splenic culture supernatant were increased, the eosinophils and goblet cells in nasal mucosa were significantly increased. The expression of encoding IL-10, TGF-β gene and eosinophils activation gene in nasal mucosa were up-regulated. As the duration of nasal dripping increased, the allergic symptoms gradually decreased, serum OVA-specific antibodies IgE, IgG1, IgG2 a continued to increase. Splenic culture supernatant IL-4 decreased, IL-10 increased, IFN-γ had a downward trend. In nasal mucosa, goblet cells decreased significantly. Genes involved in eosinophils activation were significantly down-regulated. The encoding tolerance genes such as IL-10 and TGF-β genes remained highly expressed. Ten core genes associated with immune tolerance in localized allergic rhinitis were screened, Rps27a, Uba52, Kng2, Gnal, C3, Rtp4, Reep1, Rtp2, Rtp1, Reep5. Conclusion: The mice first develop an allergy and then develop immune tolerance by continuous local drops of the allergen. The generation of immune tolerance may be induced by the continuous action of allergens, which induced systemic and local immune tolerance effects.