Summary
Background
Fraxinus excelsior (Ash) is a common tree and is important cause of winter–spring pollinosis in many temperate regions in the world. In this study, a high molecular weight allergen from ash pollen was identified.
Methods
In all, 20 individuals allergic to ash participated in the study. Characterization and immunoreactivity of ash pollen proteins was performed using sodium dodecyl sulfate polyacrylamide electrophoresis (SDS-PAGE), two-dimensional (2D) gel electrophoresis and immunoblotting.
Results
Immunoglobulin E (IgE)-binding proteins with apparent molecular mass ranging from 9 to 110 kDa were detected in ash pollen extract. Serum IgE of 7 (35%) patients reacted with a 53-kDa protein band. Analysis of 2D immunoblots showed several IgE-binding proteins. Moreover, mass spectrometry analysis indicated that the 53-kDa allergen was homologous to calreticulin.
Discussion
We defined a novel allergen from F. excelsior pollen with a molecular weight of about 53 kDa. This allergen could be considered as an important high molecular weight allergenic protein for further studies on cross-reactivity and development of diagnostic and therapeutic approaches.
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Abbreviations
- 2‑DE:
-
Two-dimensional electrophoresis
- BSA:
-
Bovine serum albumin
- CBB:
-
Comassie brilliant blue
- CRT:
-
Calreticulin
- ER:
-
Endoplasmic reticulum
- HMW:
-
High molecular weight
- IAARI:
-
Immunology, Asthma and Allergy Research Institute
- IEF:
-
Isoelectric focusing
- IPG:
-
Immobilized pH gradient
- IPI:
-
International protein index
- LiquidDAB+:
-
High-sensitivity liquid diaminobenzidine
- MALDI:
-
Matrix assisted laserdesorption/ionization
- MW:
-
Molecular weight
- PVDF:
-
Polyvinylidene fluoride
- SDS-PAGE:
-
SDS-polyacrylamide gel electrophoresis
- TBS:
-
Tris-buffered saline
- TOF:
-
Time of flight
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Acknowledgements
This study was supported by Immunology, Asthma and Allergy Research Institute (IAARI), Children’s Medical Center, Tehran University of Medical Sciences (grant # 93-03-40-27007).
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M. Sharif Shoushtari, A. Majd, M.-A. Assarehzadegan, S. Fanuel, M. Moin, T. Nejadsattari, R. S. Shoormasti,M. Badalzadeh, S. Tajik, M.R. Fazlollahi, B. Tayebi, Z. Pourpak and G.A. Kardar declare that they have no competing interests.
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Sharif Shoushtari, M., Majd, A., Assarehzadegan, MA. et al. Identification of a 53 kDa protein, as a new high molecular weight allergen from Fraxinusexcelsior (Ash) pollen. Allergo J Int 29, 233–239 (2020). https://doi.org/10.1007/s40629-020-00129-3
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DOI: https://doi.org/10.1007/s40629-020-00129-3