尘螨变应原基因修饰的树突状细胞治疗变应性鼻炎小鼠的实验研究

Experimental study on dendritic cells modified by dust mite allergen gene in the treatment of allergic rhinitis in mouse

目的探讨尘螨变应原（Der p1）基因修饰的树突状细胞（dendritic cells，DC）对变应性鼻炎（AR）小鼠的治疗效果。方法制备Der p1基因修饰的DC（Der p1－DC）。将24只健康雄性Balb／c小鼠采用随机数表法分为4组：未转染基因DC（immature DC，imDC）/AR组；糖皮质激素/AR组；Der p1－DC/AR组；imDC/正常组，每组6只。以Der p1建立AR小鼠模型，分别给予腹腔注射Der p1－DC、imDC及地塞米松，同量生理盐水处理正常对照小鼠，使用酶联免疫吸附试验（ELISA）法测定血IgE、IgG1与组胺水平；反转录聚合酶链反应（RT－PCR）与Western blot法测定小鼠鼻黏膜白细胞介素（IL）－4、IL－13的mRNA与蛋白的表达。全部数据均经SPSS 19.0软件进行统计学分析，多组均数样本比较采用方差分析。结果Der p1－DC/AR组的血IgE、IgG1与组胺水平低于imDC/AR组[（0.560±0.110）OD 450 nm比（1.150±0.280）OD 450 nm，（0.690±0.054）OD 450 nm比（0.920±0.125）OD 450 nm，（4 145±670）pg/ml比（7 685±669）pg/ml，t值分别为4.80、4.14、9.16，P值均〈0.05]，鼻黏膜中IL－4与IL－13的表达明显低于imDC/AR组（0.41±0.25比1.59±1.02，0.26±0.01比1.10±0.09，t值分别为2.75、22.72，P值均〈0.05），与地塞米松治疗组相比差异无统计学意义。结论Der p1－DC可以减轻AR小鼠的炎症，并降低IL－4与IL－13的表达，提示其可用于AR的免疫治疗。

ObjectiveTo explore the therapeutic effects of dendritic cells （DC） modified by the dust-mite-allergen（Der p1） gene on mouse model of allergic rhinitis （AR）.MethodsDC modified by the Der p1 gene （Der p1-DC） were prepared.Using random number table, 24 Balb／c mice were divided into four groups： immature DC （imDC）/AR group, dexamethasone/AR group, Der p1-DC/AR group and control group, with 6 mice in each group.AR mouse model was built with Der p1 and the mouse model of AR was established.The AR mice were respectively given by abdominal injection of Der p1-DC, imDC and dexamethasone.Normal control mice were treated with physiologic saline.ELISA method was used for determining the content of IgE, IgG1and histamine in blood.The relative expression of mRNA of IL-4 and IL-13 on nasal mucosa with protein was analyzed by RT-PCR and Westen blot methods.All the data were statistically analyzed by SPSS 19.0 statistical software, and the variance analysis was used in multiple groups of average samples.ResultsThe contents of IgE, IgG1 and histamine in the mice of Der p1-DC/AR group were lower than those in imDC/AR group （（0.560±0.110） OD 450 nm vs （1.150±0.280） OD 450 nm, （0.690±0.054） OD 450 nm vs （0.920±0.125） OD 450 nm, （4 145±670） pg/ml vs （7 685±669） pg/ml, t value was 4.80, 4.14, 9.16, respectively, all P〈0.05）, and the expression of IL-4 and IL-13 on nasal mucosa in Der p1-DC/AR group was remarkedly lower than those in imDC/AR group （0.41±0.25 vs 1.59±1.02, 0.26±0.01 vs 1.10±0.09, t value was 2.75, 22.72, respectively, all P〈0.05）. There was no statistically significant difference between the mice treated with Der p1-DC and dexamethasone group.ConclusionsThe results showed that Der p1-DC could reduce inflammation in AR mice and decrease the expression of IL-4 and IL-13. It suggested that Der p1-DC can be used in the immunotherapy of AR mouse.