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Molecular and Immunochemical Characterization of Pop N 2: A New Allergen of Populus Nigra Pollen Publisher Pubmed



Shams MH1, 2 ; Assarehzadegan MA3, 4 ; Eskandari N1 ; Masjedi M1 ; Kheirandish F2, 5 ; Ghasemi R1 ; Ganjalikhani Hakemi M1 ; Varzi AM6 ; Safari M7 ; Sohrabi SM2 ; Abdoli Sereshki H3, 4
Authors
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Authors Affiliations
  1. 1. Department of Immunology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
  2. 2. Razi Herbal Medicines Research Center, Lorestan University of Medical sciences, Khorramabad, Iran
  3. 3. Immunology Research Center, Institute of Immunology and Infectious Diseases, Iran University of Medical Science, Tehran, Iran
  4. 4. Immunology Department, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
  5. 5. Department of Medical Parasitology and Mycology, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran
  6. 6. Department of Medical Immunology, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran
  7. 7. Department of Pediatrics, School of Medicines, Hamedan University of Medical Science, Hamedan, Iran

Source: Clinical and Experimental Allergy Published:2021


Abstract

Background: Pollen is one of the most common allergens that cause respiratory allergies worldwide. Pollen grains from poplars have been reported as important sources of pollinosis in many countries. Objective: The aim of the present study was to determine the molecular and immunochemical characterization of Pop n 2, a novel allergen of Populus nigra (P nigra) pollen extract. Methods: In this study, the pollen extract of P nigra was analysed by SDS-PAGE, and the allergenic profile was determined by IgE immunoblotting and specific ELISA using the sera of twenty allergic patients. The coding sequence of Pop n 2 was cloned and expressed in the Escherichia coli BL21 (DE3) using plasmid the pET-21b (+). Finally, the expressed recombinant Pop n 2 was purified by affinity chromatography. Results: Pop n 2 belongs to the profilin family with a molecular weight of approximately 14 kDa. Pop n 2 is the most IgE-reactive protein (about 65%) in the P nigra pollen extract. The cDNA sequencing results indicated an open reading frame 396 bp that encodes 131 amino acid residues. The results of ELISA and Immunoblotting assays showed that recombinant Pop n 2 could react with the IgE antibody in patients' sera, like its natural counterpart. Conclusion: Our data revealed that Pop n 2 is a significant allergen in the P nigra pollen extract. Moreover, we observed that the recombinant Pop n 2 produced by the pET-21b (+) vector in the E colisystem acts as its natural counterpart. © 2021 John Wiley & Sons Ltd
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Experts (# of related papers)
Nahid Eskandari (1)