Expression and characterization of recombinant Par j 1 and Par j 2 resembling the allergenic epitopes of Parietaria judaica pollen

• Published in: Scientific reports 2019-10, Vol.9 (1), p.15043-12, Article 15043
• Main Authors: Dorofeeva, Yulia, Colombo, Paolo, Blanca, Miguel, Mari, Adriano, Khanferyan, Roman, Valenta, Rudolf, Focke-Tejkl, Margarete
• Format: Article
• Language: English
• Subjects: 631/250; 631/250/256; 82; 82/1; 82/58; 82/80; 82/83; Adolescent; Adult; Allergens; Allergens - metabolism; Allergies; Amino Acid Sequence; Antigens, Plant - metabolism; Asthma; Biophysical Phenomena; Cell Line; Child; Circular dichroism; Cross Reactions; E coli; Enzyme-linked immunosorbent assay; Epitopes; Epitopes - chemistry; Epitopes - metabolism; Escherichia coli - metabolism; Female; Hay fever; Humanities and Social Sciences; Humans; Hypersensitivity - immunology; Immunization; Immunoglobulin E; Immunoglobulin E - immunology; Immunoglobulin G; Immunotherapy; Insect cells; Insects; Judaica; Lipid transfer proteins; Male; Middle Aged; multidisciplinary; Parietaria; Parietaria judaica; Plant Extracts - metabolism; Plant Proteins - chemistry; Plant Proteins - isolation & purification; Plant Proteins - metabolism; Pollen; Recombinant Proteins - chemistry; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Science; Science (multidisciplinary); Young Adult
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-019-50854-1

The weed wall pellitory, Parietaria judaica , is one the most important pollen allergen sources in the Mediterranean area causing severe symptoms of hay fever and asthma in allergic patients. We report the expression of the major Parietaria allergens, Par j 1 and Par j 2 which belong to the family of lipid transfer proteins, in insect cells. According to circular dichroism analysis and gel filtration, the purified allergens represented folded and monomeric proteins. Insect cell-expressed, folded Par j 2 exhibited higher IgE binding capacity and more than 100-fold higher allergenic activity than unfolded Escherichia coli -expressed Par j 2 as demonstrated by IgE ELISA and basophil activation testing. IgE ELISA inhibition assays showed that Par j 1 and Par j 2, contain genuine and cross-reactive IgE epitopes. IgG antibodies induced by immunization with Par j 2 inhibited binding of allergic patients IgE to Par j 1 only partially. IgE inhibition experiments demonstrated that insect cell-expressed Par j 1 and Par j 2 together resembled the majority of allergenic epitopes of the Parietaria allergome and therefore both should be used for molecular diagnosis and the design of vaccines for allergen-specific immunotherapy of Parietaria allergy.