Introduction and objectives

Allergen-specific immunotherapy (ASIT) is the only allergic disease-modifying therapy available for children and adults, and recombinant allergens are an interesting approach to improve allergy diagnosis and ASIT. Tyrophagus putrescentiae is a common storage mite that produces potent allergens. The aim of this study was to express and characterize recombinant group 4 allergen protein of T. putrescentiae (Tyr p 4), and to further investigate allergenicity and potential epitopes of Tyr p 4.

Materials and methods

The cDNA encoding Tyr p 4 was generated by RT-PCR and subcloned into pET-28a(+) plasmid. The plasmid was then transformed into E. coli cells for expression. After purification by nickel affinity chromatography and identification by SDS-PAGE, recombinant Tyr p 4 protein was used for a skin prick test and an ELISA to determine the allergic response.

Results

Study participants’ allergic response rate to Tyr p 4 protein was 13.3% (16/120). Eight B-cell epitopes and three T-cell epitopes of Tyr p 4 were predicted.

Conclusions

Similar to group 4 allergens of other species of mite, allergenicity of Tyr p 4 is weak. The expression, characterization and epitope prediction of recombinant Tyr p 4 protein provide a foundation for further study of this allergen in the diagnosis and ASIT of storage mite allergy.

中文翻译：

---

Tyrophagus putrescentiae 组 4 变应原变应原性和表位预测。

介绍和目标

过敏原特异性免疫疗法 (ASIT) 是唯一可用于儿童和成人的过敏性疾病改善疗法，而重组过敏原是改善过敏诊断和 ASIT 的一种有趣方法。Tyrophagus putrescentiae是一种常见的储藏螨，会产生强效过敏原。本研究的目的是表达和表征腐烂木霉重组体第 4 组过敏原蛋白(Tyr p 4)，并进一步研究 Tyr p 4 的致敏性和潜在表位。

材料和方法

编码 Tyr p 4 的 cDNA 通过 RT-PCR 产生并亚克隆到 pET-28a(+) 质粒中。然后将质粒转化到大肠杆菌细胞中进行表达。经镍亲和层析纯化和SDS-PAGE鉴定后，重组Tyr p 4蛋白用于皮肤点刺试验和ELISA确定过敏反应。

结果

研究参与者对 Tyr p 4 蛋白的过敏反应率为 13.3% (16/120)。预测了 Tyr p 4 的八个 B 细胞表位和三个 T 细胞表位。

结论

与其他螨类的第 4 组过敏原相似，Tyr p 4 的致敏性较弱。重组Tyr p 4蛋白的表达、表征和表位预测为进一步研究该变应原在储螨变态反应的诊断和ASIT中的应用奠定了基础。