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[屋尘螨变应原2 T细胞表位疫苗对哮喘小鼠的特异性免疫治疗作用]

[A specific immune therapeutic effect of Der p2 T cell epitope vaccine on asthma mice].

作者信息

Xiao-Dong Zhan, Bin-Bin Duan, Yong Hong, Chao-Pin Li

机构信息

Anhui Province Key Laboratory of Active Biological Macro-molecules Research, Wannan Medical University, Wuhu 241002, China.

出版信息

Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi. 2016 Dec 12;29(1):59-63. doi: 10.16250/j.32.1374.2016120.

Abstract

OBJECTIVE

To investigate the specific immune therapeutic effect of the T cell fusion peptide vaccine from group II allergens from (Der p2).

METHODS

Thirty mice were randomly divided into three groups, namely a negative control group (a PBS group), positive control group (an asthma group) and protein Der p2 T cell fusion epitope for specific immunotherapy (SIT) group (a Der p2 T group). The extract of house dust mites (HDM) was used to establish the asthmatic models in BALB/c mice, and the PBS group was always used with PBS buffer. Thirty minutes before spray inhalation from 25 to 27 days, the mice of the Der p2 T group were respectively injected subcutaneously with the therapeutic proteins for SIT, then the serum and bronchoalveolar lavage fluid (BALF) were collected. ELISA was used to assay the levels of IFN-γ, IL-4, and IL-13 in BALF, as well as serum levels of specific IgE and IgG2a. The lung tissue sections were stained with haematoxylin and eosin (H&E) for pathological examinations.

RESULTS

The ELISA detection revealed that the number of eosinophil in BALF of the asthmatic mice was (5.57 ± 0.64)×10/ml, which was significantly higher than that in the PBS group [(0.50 ±0.30)×10/ml, < 0.01], the number of eosinophil in the Der p2 T immunotherapy group decreased significantly [(3.45 ± 0.36)×10/ml, < 0.01]. The content of IFN-γ in the PBS group, asthma group and Der p2 T group were (267.00 ± 21.98), (155.80 ± 20.53) pg/ml and (234.40 ± 24.46) pg/ml respectively. Compared with the asthma group, the mice with Der p2 T vaccine specific immune treatment produced a high level of IFN-γ ( < 0.01). The content of IL-4 in the PBS group, asthma group and Der p2 T group were (23.40 ± 5.96), (53.28 ± 8.26) pg/ml and (30.00 ± 5.50) pg/ml respectively. Compared with the asthma group, the content of IL-4 in the mice of the Der p2 T treatment group was significantly lower ( < 0.01). Compared with the asthma group [(308.10 ± 28.32) pg/ml], the content of IL-13 in BALF of the mice in the Der p2 T treatment group was significantly decreased, which was [(174.50 ± 25.99) pg/ml, < 0.01]. The content of IL-13 in the PBS group was (95.99 ± 31.14) pg/ml. The lung tissue sections showed that the lung inflammation in the p2 T Der group was significantly less than that in the asthma group, and the inflammatory cell infiltration was significantly decreased, and airway epithelial construction remodeled.

CONCLUSIONS

The Der p2 T cell fusion epitope, which is as vaccines for specific immunotherapy with asthma models, can alleviate effectively allergic inflammation of airway and lung in the mice, and it may be used as a candidate vaccine for asthma.

摘要

目的

探讨来自粉尘螨(Der p2)Ⅱ组变应原的T细胞融合肽疫苗的特异性免疫治疗效果。

方法

将30只小鼠随机分为3组,即阴性对照组(PBS组)、阳性对照组(哮喘组)和用于特异性免疫治疗(SIT)的蛋白Der p2 T细胞融合表位组(Der p2 T组)。用屋尘螨(HDM)提取物在BALB/c小鼠中建立哮喘模型,PBS组始终使用PBS缓冲液。在第25至27天喷雾吸入前30分钟,Der p2 T组小鼠分别皮下注射用于SIT的治疗性蛋白,然后收集血清和支气管肺泡灌洗液(BALF)。采用ELISA法检测BALF中IFN-γ、IL-4和IL-13的水平,以及血清中特异性IgE和IgG2a的水平。肺组织切片用苏木精和伊红(H&E)染色进行病理检查。

结果

ELISA检测显示,哮喘小鼠BALF中嗜酸性粒细胞数量为(5.57±0.64)×10/ml,显著高于PBS组[(0.50±0.30)×10/ml,P<0.01],Der p2 T免疫治疗组嗜酸性粒细胞数量显著减少[(3.45±0.36)×10/ml,P<0.01]。PBS组、哮喘组和Der p2 T组中IFN-γ的含量分别为(267.00±21.98)、(155.80±20.53)pg/ml和(234.40±24.46)pg/ml。与哮喘组相比,接受Der p2 T疫苗特异性免疫治疗的小鼠产生了高水平的IFN-γ(P<0.01)。PBS组、哮喘组和Der p2 T组中IL-4的含量分别为(23.40±5.96)、(53.28±8.26)pg/ml和(30.00±5.50)pg/ml。与哮喘组相比,Der p2 T治疗组小鼠中IL-4的含量显著降低(P<0.01)。与哮喘组[(308.10±28.32)pg/ml]相比,Der p2 T治疗组小鼠BALF中IL-13的含量显著降低,为[(174.50±25.99)pg/ml,P<0.01]。PBS组中IL-13的含量为(95.99±31.14)pg/ml。肺组织切片显示,Der p2 T组的肺部炎症明显少于哮喘组,炎症细胞浸润显著减少,气道上皮结构重塑。

结论

Der p2 T细胞融合表位作为哮喘模型特异性免疫治疗的疫苗,可有效减轻小鼠气道和肺部的过敏性炎症,可能作为哮喘的候选疫苗。

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