Foxp3+Treg cells mediate immune protection of humulus pollen allergy DNA vaccine pcD-NA3.1-Hum in asthmatic mice
- VernacularTitle:葎草花粉变应原核酸疫苗通过诱导Foxp3+Treg细胞分化介导对哮喘模型小鼠的免疫保护作用
- Author:
Jiamei LU
1
;
Manxiang LI
;
Xiuzhen SUN
;
Yonghong ZHANG
;
Yun LIU
;
Jing XU
;
Sumei ZHANG
Author Information
1. 西安交通大学医学院第二附属医院呼吸病研究室
- Keywords:
allergic asthma;
humulus pollen allergy;
DNA vaccine;
immunogenicity;
Foxp3+Treg cells
- From:
Journal of Southern Medical University
2014;(1):14-19
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a humulus pollen allergy DNA vaccine pcDNA3.1-Hum and investigate its effect for immune protection mediated by Foxp3+Treg cells in asthmatic mice. Methods The target humulus gene obtained from pTripIEx2-Hum plasmid by double enzyme digestion was inserted sequentially into pcDNA3.1(-) vector to generate the recombinant plasmid pcDNA3.1-Hum, which was validated by sequencing. The pcDNA3.1-Hum plasmid was transfected into COS-7 cells and the expression of the ectopic protein was analyzed using Western blotting. Co-cultured dendritic cells and CD4+CD25-T cells were stimulated with the expressed protein to test its efficacy in inducing Foxp3+Treg cells. The levels of humulus-specific IgE and IgG2a were assayed to evaluate the allergenicity and immunogenicity of pcDNA3.1-Hum in mice. The immunoprotective effect of pcDNA3.1-Hum was assessed in a mouse model of humulus-specific asthma. Results The constructed pcDNA3.1-Hum plasmid was validated by sequencing and Western blotting, and the expressed protein was shown to induce Foxp3+Treg cells in the co-culture. In normal mice, pcDNA3.1-Hum induced a significant increase of humulus-specific IgG2a but had no effect on IgE. In the asthmatic mice, pcDNA3.1-Hum significantly decreased inflammatory cell counts and eosinophil percentages in the BALF, ameliorated lung inflammation, and lowered AHR and IL-4 levels; immunization of the mice with pcDNA3.1-Hum reversed humulus-induced reduction of serum IFN-γ and prevented the humulus-triggered reduction of Foxp3+Treg cell percentage in the spleen. Conclusion We have successfully constructed a highly immunogenic pcDNA3.1-Hum DNA vaccine that can mediate immune protection by inducing Foxp3+Treg cells.
