Expression and characterization of recombinant Par j 1 and Par j 2 resembling the allergenic epitopes of Parietaria judaica pollen
Abstract 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...
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| Format: | Article |
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Nature Portfolio
2019-10-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-019-50854-1 |
| _version_ | 1818996557153828864 |
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| author | Yulia Dorofeeva Paolo Colombo Miguel Blanca Adriano Mari Roman Khanferyan Rudolf Valenta Margarete Focke-Tejkl |
| author_facet | Yulia Dorofeeva Paolo Colombo Miguel Blanca Adriano Mari Roman Khanferyan Rudolf Valenta Margarete Focke-Tejkl |
| author_sort | Yulia Dorofeeva |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-20T21:31:38Z |
| format | Article |
| id | doaj.art-a736b1f6001d4b09a56aa18c1c6b4494 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-20T21:31:38Z |
| publishDate | 2019-10-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-a736b1f6001d4b09a56aa18c1c6b44942022-12-21T19:26:03ZengNature PortfolioScientific Reports2045-23222019-10-019111210.1038/s41598-019-50854-1Expression and characterization of recombinant Par j 1 and Par j 2 resembling the allergenic epitopes of Parietaria judaica pollenYulia Dorofeeva0Paolo Colombo1Miguel Blanca2Adriano Mari3Roman Khanferyan4Rudolf Valenta5Margarete Focke-Tejkl6Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology and Immunology, Medical University of ViennaIstituto di Biomedicina ed Immunologia Molecolare “Alberto Monroy” del Consiglio Nazionale delle RicercheHospital CivilAssociated Centers for Molecular AllergologyRussian People’s Friendship UniversityDivision of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology and Immunology, Medical University of ViennaDivision of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology and Immunology, Medical University of ViennaAbstract 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.https://doi.org/10.1038/s41598-019-50854-1 |
| spellingShingle | Yulia Dorofeeva Paolo Colombo Miguel Blanca Adriano Mari Roman Khanferyan Rudolf Valenta Margarete Focke-Tejkl Expression and characterization of recombinant Par j 1 and Par j 2 resembling the allergenic epitopes of Parietaria judaica pollen Scientific Reports |
| title | Expression and characterization of recombinant Par j 1 and Par j 2 resembling the allergenic epitopes of Parietaria judaica pollen |
| title_full | Expression and characterization of recombinant Par j 1 and Par j 2 resembling the allergenic epitopes of Parietaria judaica pollen |
| title_fullStr | Expression and characterization of recombinant Par j 1 and Par j 2 resembling the allergenic epitopes of Parietaria judaica pollen |
| title_full_unstemmed | Expression and characterization of recombinant Par j 1 and Par j 2 resembling the allergenic epitopes of Parietaria judaica pollen |
| title_short | Expression and characterization of recombinant Par j 1 and Par j 2 resembling the allergenic epitopes of Parietaria judaica pollen |
| title_sort | expression and characterization of recombinant par j 1 and par j 2 resembling the allergenic epitopes of parietaria judaica pollen |
| url | https://doi.org/10.1038/s41598-019-50854-1 |
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