Dermatophagoides pteronyssinus allergen Der p 22: Cloning, expression, IgE‐binding in asthmatic children, and immunogenicity

免疫原性 医学 过敏原 免疫学 免疫球蛋白E 屋尘螨 过敏 草甘膦科 重组DNA 细胞因子 过敏性炎症 抗原 抗体 生物 基因 生物化学
作者
Ying Zhou,Lin Li,Zhiwei Yu,Xue‐Ping Gu,Ruilin Pan,Qingqing Li,Cunyin Yuan,Fangfang Cai,Yimin Zhu,Yubao Cui
出处
期刊:Pediatric Allergy and Immunology [Wiley]
卷期号:33 (8) 被引量:17
标识
DOI:10.1111/pai.13835
摘要

Abstract Background Dust mite extract contains multiple components that, while useful in clinical allergy diagnosis and treatment, can cause serious side effects. Defining components of dust mite extract is important their contributions to allergic disease. This study aimed to characterize a novel dust mite allergen, Der p 22. Methods We amplified the cDNA encoding Der p 22 from total RNA of the mite Dermatophagoides pteronyssinus, and inserted it into an expression construct for transformation to competent cells. Purified recombinant (r) Der p 22 was tested for IgE‐binding reactivity in sera obtained from children with allergic asthma by the Affiliated Wuxi Children's Hospital of Nanjing Medical University (Jiangsu, China). rDer p 22 also was used to challenge BALB/c mice to assess effects on T helper cells and cytokine levels and applied to cultured lung epithelial cells to evaluate apoptosis and cytokine secretion. Results rDer p 22 bound to IgE in 93.75% of sera from pediatric allergic asthma patients. Mice challenged with rDer p 22 had altered Th1/Th2 ratios in spleen and lymph, and lower levels of cytokines IFN‐γ but higher levels of IL‐4 and IL‐10 in alveolar lavage fluid compared with controls ( p < .05). Cultured lung epithelial cells had greater apoptosis rates and exhibited higher levels of IL‐6, IL‐8, and GM‐CSF when treated with rDer p 22 compared with control treatment ( p < .05). Conclusions Recombinant Der p 22 exhibited high IgE‐binding rates in allergic children, indicating the activity of the recombinant protein and suggesting this novel allergen may be appropriate for inclusion in an allergy diagnostic workup. This finding is supported by in vitro and mouse in vivo studies showing rDer p 22 induced strong allergenic reactivity and apoptosis.
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