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重组屋尘螨第2组变应原疫苗免疫治疗抑制小鼠过敏性气道炎症

[Immunotherapy with recombinant house dust mite group 2 allergen vaccine inhibits allergic airway inflammation in mice].

作者信息

Yu Hai-Qiong, Liu Zhi-Gang, Yu Kun-Ying, Xu Zuo-Qian, Qiu Jing

机构信息

Allergy and Immunology Institute , Shenzhen University, Shenzhen 518060, China.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2006 Dec;24(6):414-9.

Abstract

OBJECTIVE

To investigate the efficacy and mechanism of subcutaneously given recombinant Der p 2 entrapped PLGA nanoparticles (DEPN) on mouse model with allergic airway inflammation.

METHODS

40 BALB/c mice were randomly divided into 5 groups, group A (normal control) were treated with saline (100 microl) all the time, groups B, C, D and E were sensitized intraperitoneally with crude dust mite extracts (10 microg) and then subcutaneously treated respectively with PBS (100 microl), 2 mg empty PLGA (EP), 100 microg rDerp2, and 2 mg DEPN (loaded with 100 microg rDerp 2) for 3 times, once per day, followed by intranasal challenge of 50 microg rDer p 2. One day post challenge, mice were sacrificed and bronchoalveolar lavage fluid (BALF) was collected. Number of the total cells and eosinophils was determined, and airway inflammation and mucus secretion were analyzed by haematoxylin and eosin (H&E) staining and periodic acid-Schiff (PAS) staining. Level of cytokines in the supernatant of splenocyte culture was assayed by ELISA. Level of rDer p 2 specific IgG2a and IgE in the sera was determined by ELISA.

RESULTS

The lung histology showed development of eosinophil infiltration in the airway of mice in groups B and C. The lung inflammation and mucus secretion in groups D and E were significantly alleviated than that of groups B and C. Number of total cells (63.50+/-5.12) and eosinophils (15.32+/-3.04) in BALF decreased in group B. Compared with group B, the number of total cells in groups D (55.3+/-5.20) x 10(4) /ml and E (41.00+/-4.91) x 10(4)/ml greatly decreased (P<0.05), and same with that of eosinophils in groups D (9.56+/-1.09) x 10(4) /ml and E (3.22+/-0.31) x 10(4)/ml. The rDerp 2 specific IgE and IgG2a antibodies in group B were 1.14+/-0.105 and 0.14+/-0.07 respectively. The level of specific IgE was significantly lower (P<0.01) in groups D (0.93+/-0.04) and E (0.77+/-0.09), and that of IgG2a in groups D (1.02+/-0.01) and E (1.17+/-0.46) were significantly higher (P<0.01) than that in group B. The level of IL-4 and IFN-gamma in BALF in group B were (78.90+/-6.07) pg/ml and (27.30+/-3.51) pg/ml respectively. IL-4 in groups D and E was (55.6+/-3.79) pg/ml and (48.6+/-4.50)pg/ml respectively, significantly lower (P<0.01) than that of group B; while IFN-gamma (68.50+/-2.87) pg/ml in group E was significantly higher than that of group B (P<0.01). IL-4 released from cultured splenocytes in groups D and E was (56.30+/-4.85) pg/ml and (40.20+/-4.36) pg/ml respectively, significantly lower than that in group B (81.2+/-6.84 pg/ml) (P<0.01). The released IFN-gamma in group E was (70.20+/-3.85) pg/ml, significantly higher than in group B (34.60+/-2.25) pg/ml (P<0.01).

CONCLUSION

DEPN can inhibit airway allergic inflammation, its mechanism may be relevant to a balance of Th1 and Th2.

摘要

目的

探讨皮下注射包裹重组Der p 2的聚乳酸-羟基乙酸共聚物纳米粒(DEPN)对小鼠变应性气道炎症模型的疗效及作用机制。

方法

将40只BALB/c小鼠随机分为5组,A组(正常对照组)始终给予生理盐水(100 μl),B、C、D、E组小鼠腹腔注射粗制尘螨提取物(10 μg)致敏,然后分别皮下注射PBS(100 μl)、2 mg空白聚乳酸-羟基乙酸共聚物(EP)、100 μg重组Der p 2、2 mg DEPN(负载100 μg重组Der p 2),每日1次,共3次,随后经鼻激发50 μg重组Der p 2。激发后1天,处死小鼠,收集支气管肺泡灌洗液(BALF)。测定总细胞数和嗜酸性粒细胞数,采用苏木精-伊红(H&E)染色和过碘酸-希夫(PAS)染色分析气道炎症和黏液分泌情况。采用酶联免疫吸附测定(ELISA)法检测脾细胞培养上清液中细胞因子水平。采用ELISA法测定血清中重组Der p 2特异性IgG2a和IgE水平。

结果

肺组织学检查显示,B组和C组小鼠气道出现嗜酸性粒细胞浸润。D组和E组的肺部炎症和黏液分泌较B组和C组明显减轻。B组BALF中总细胞数(63.50±5.12)和嗜酸性粒细胞数(15.32±3.04)减少。与B组相比,D组(55.3±5.20)×10⁴/ml和E组(41.00±4.91)×10⁴/ml的总细胞数显著减少(P<0.05),D组(9.56±1.09)×10⁴/ml和E组(3.22±0.31)×10⁴/ml的嗜酸性粒细胞数也显著减少。B组重组Der p 2特异性IgE和IgG2a抗体分别为1.14±0.105和0.14±0.07。D组(0.93±0.04)和E组(0.77±0.09)的特异性IgE水平显著降低(P<0.01),D组(1.02±0.01)和E组(1.17±0.46)的IgG2a水平显著高于B组(P<0.01)。B组BALF中IL-4和IFN-γ水平分别为(78.90±6.07)pg/ml和(27.30±3.51)pg/ml。D组和E组的IL-4分别为(55.6±3.79)pg/ml和(48.6±4.50)pg/ml,显著低于B组(P<0.01);而E组的IFN-γ(68.50±2.87)pg/ml显著高于B组(P<0.01)。D组和E组培养的脾细胞释放的IL-4分别为(56.30±4.85)pg/ml和(40.20±4.36)pg/ml,显著低于B组(81.2±6.84 pg/ml)(P<0.01)。E组释放的IFN-γ为(70.20±3.85)pg/ml,显著高于B组(34.60±2.25)pg/ml(P<0.01)。

结论

DEPN可抑制气道变应性炎症,其机制可能与Th1和Th2平衡有关。

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