Expression, epitope prediction and IgE‑binding of the <em>Tyrophagus putrescentiae</em> group 13 allergen

Wang,Nan; Zhou,Ying; Wu,Meili; Zhu,Hanting; Cui,Yubao

doi:10.3892/etm.2020.9149

Expression, epitope prediction and IgE‑binding of the Tyrophagus putrescentiae group 13 allergen

Authors:
- Nan Wang
- Ying Zhou
- Meili Wu
- Hanting Zhu
- Yubao Cui
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Affiliations: Department of Basic Medicine, Jiangsu Vocational College of Medicine, Yancheng, Jiangsu 224005, P.R. China, Department of Pediatrics Laboratory, Wuxi Children's Hospital, Wuxi, Jiangsu 214023, P.R. China, Department of Clinical Laboratory, Wuxi People's Hospital Affiliated to Nanjing Medical University, Wuxi, Jiangsu 214023, P.R. China
Published online on: August 27, 2020 https://doi.org/10.3892/etm.2020.9149
Article Number: 21
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Storage mites, such as Tyrophagus putrescentiae, are an important source of allergens that cause allergic diseases in humans. It has previously been indicated that T. putrescentiae has a high sensitization rate as an allergen in some Asian and European countries. Identifying and cloning the allergens in this species may enable improved diagnostic and therapeutic approaches. The aim of the present study was to clone and sequence the T. putrescentiae group 13 allergen (Tyr p 13) isolated from storage mites in China, to use bioinformatics tools to model its biophysical characteristics and to induce protein expression to test its IgE‑binding activity. The full‑length cDNA comprised 486 bp and was predicted to include a signal peptide of 22 amino acids. Its secondary structure was shown to comprise an α‑helix (10.79%), extended strand (33.81%) and random coils (55.40%). Using homology modeling, the present study constructed a reasonable tertiary structure of Tyr p 13. Linear Bcell epitopes at amino acids 47‑53, 70‑76, 81‑86, 101‑105 and 112120 were predicted. Three discontinuous B‑cell epitopes were also predicted: i) 47, 48, 49, 50, 51, 52, 53, 70, 71, 72 and 73; ii) 91, 92, 93, 94, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121 and 138; and iii) 74, 76, 79, 81, 82, 83, 84, 86, 101, 102, 103, 104 and 105. SDS‑PAGE identified a specific band at the predicted molecular weight of the recombinant Tyr p 13 (rTyr p 13), demonstrating its successful expression. The rTyr p 13 bound to IgE in the serum of 13.2% (5/38) of patients allergic to T. putrescentiae, according to ELISA. The successful cloning of Tyr p 13 and basic bioinformatics analysis of the protein provided a foundation for the further study of this allergen with regards to the diagnosis and treatment of patients allergic to storage mites. These results provided a theoretical basis for the design of rTyr p 13 with modified B‑cell epitopes.

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