Zhong-Shan Gao1,2, Xiang Zhou1, Zhao-Wei Yang3, Serge A Versteeg4, Ling Gao1, Wan-Yi Fu1, Hui-Ying Wang5, Jian-Ying Zhou6, Jaap H Akkerdaas4, Ronald van Ree4,7. 1. Department of Horticulture, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China. 2. Allergy Research Center, Zhejiang University, Hangzhou, China. 3. State Key Laboratory of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, China. 4. Department of Experimental Immunology, Academic Medical Center-University of Amsterdam, Amsterdam, The Netherlands. 5. Department of Allergy, The Second Affiliated Hospital of Zhejiang University, School of Medicine, Hangzhou, China. 6. Department of Allergy, The First Affiliated Hospital of Zhejiang University, School of Medicine, Hangzhou, China. 7. Department of Otorhinolaryngology, Academic Medical Center-University of Amsterdam, Amsterdam, The Netherlands.
Abstract
SCOPE: Pru p 1, the Bet v 1 homologue from peach, has been identified as a clinically relevant allergen. Three isoforms have been described, two in peach fruit (Pru p 1.0101 and Pru p 1.0201) and one in pollen (Pru p 1.0301). The present study aimed to compare their IgE-binding potencies. METHODS AND RESULTS: Three Pru p 1 isoforms were cloned and expressed as soluble proteins with His-tags in Escherichia coli. Protein identity was confirmed by MS, circular dichroism, and RNAse activity. IgE-binding capacity using ELISA and ImmunoCAP was compared. Three Pru p 1 isoforms had quite similar IgE-binding potencies for 60% of the sera, but more than twofold between any two isoforms among 40% of the 47 sera. The mean IgE binding of Pru p 1.0201 was slightly higher than other two isoforms. In a sera pool, homologous ImmunoCAP inhibition was higher than other two heterologous isoforms. Individual serum with diverse IgE values of three isoforms demonstrated the higher IgE inhibition of specific isoform with higher IgE value. CONCLUSION: A similar and variable pattern of IgE recognition was observed among three Pru p 1 isoforms. The two new isoforms can be used as more accurate diagnostic reagents.
SCOPE: Pru p 1, the Bet v 1 homologue from peach, has been identified as a clinically relevant allergen. Three isoforms have been described, two in peach fruit (Pru p 1.0101 and Pru p 1.0201) and one in pollen (Pru p 1.0301). The present study aimed to compare their IgE-binding potencies. METHODS AND RESULTS: Three Pru p 1 isoforms were cloned and expressed as soluble proteins with His-tags in Escherichia coli. Protein identity was confirmed by MS, circular dichroism, and RNAse activity. IgE-binding capacity using ELISA and ImmunoCAP was compared. Three Pru p 1 isoforms had quite similar IgE-binding potencies for 60% of the sera, but more than twofold between any two isoforms among 40% of the 47 sera. The mean IgE binding of Pru p 1.0201 was slightly higher than other two isoforms. In a sera pool, homologous ImmunoCAP inhibition was higher than other two heterologous isoforms. Individual serum with diverse IgE values of three isoforms demonstrated the higher IgE inhibition of specific isoform with higher IgE value. CONCLUSION: A similar and variable pattern of IgE recognition was observed among three Pru p 1 isoforms. The two new isoforms can be used as more accurate diagnostic reagents.
Authors: Andrea Wangorsch; Stephan Scheurer; Miguel Blanca; Natalia Blanca-Lopez; María Luisa Somoza; Laura Martín-Pedraza Journal: Front Allergy Date: 2022-02-08