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

Dorofeeva, Yulia; Colombo, Piergiuseppe; Blanca, Miguel; Mari, Adriano; Khanferyan, R. A.; Valenta, Rudolf; Focke‐Tejkl, Margarete

doi:10.1038/s41598-019-50854-1

Cited by 6 publications

(5 citation statements)

References 49 publications

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“…In Middle and Southern Europe and America grass pollen allergy dominates ( 88 ) and can be approached with BM32. In the Mediterranean area olive pollen and certain weeds such Parietaria are common requiring hyposensitization with Ole e 1 and Par j 1 as well as Par j 2 ( 89 , 90 ), respectively. In Eastern Europe and parts of the USA ragweed dominates and mugwort pollen is important in Central Asia ( 91 , 92 ).…”

Section: Unique Features Of Recombinant B Cell Epitope-based Peptide mentioning

confidence: 99%

Preventive Allergen-Specific Vaccination Against Allergy: Mission Possible?

et al. 2020

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Vaccines for infectious diseases have improved the life of the human species in a tremendous manner. The principle of vaccination is to establish de novo adaptive immune response consisting of antibody and T cell responses against pathogens which should defend the vaccinated person against future challenge with the culprit pathogen. The situation is completely different for immunoglobulin E (IgE)-associated allergy, an immunologically-mediated hypersensitivity which is already characterized by increased IgE antibody levels and T cell responses against per se innocuous antigens (i.e., allergens). Thus, allergic patients suffer from a deviated hyper-immunity against allergens leading to inflammation upon allergen contact. Paradoxically, vaccination with allergens, termed allergen-specific immunotherapy (AIT), induces a counter immune response based on the production of high levels of allergen-specific IgG antibodies and alterations of the adaptive cellular response, which reduce allergen-induced symptoms of allergic inflammation. AIT was even shown to prevent the progression of mild to severe forms of allergy. Consequently, AIT can be considered as a form of therapeutic vaccination. In this article we describe a strategy and possible road map for the use of an AIT approach for prophylactic vaccination against allergy which is based on new molecular allergy vaccines. This road map includes the use of AIT for secondary preventive vaccination to stop the progression of clinically silent allergic sensitization toward symptomatic allergy and ultimately the prevention of allergic sensitization by maternal vaccination and/or early primary preventive vaccination of children. Prophylactic allergy vaccination with molecular allergy vaccines may allow halting the allergy epidemics affecting almost 30% of the population as it has been achieved for vaccination against infectious diseases.

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Section: Unique Features Of Recombinant B Cell Epitope-based Peptide mentioning

confidence: 99%

Preventive Allergen-Specific Vaccination Against Allergy: Mission Possible?

et al. 2020

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“…For this purpose, recombinant Amb a 6 was produced in a eukaryotic expression system, i.e., in insect cells. Eukaryotic expression in insect cells was chosen because previous studies have found for the nsLTP from Parietaria judaica pollen, i.e., Par j 2, that the insect cell-expressed allergen exhibited a higher IgE reactivity and allergenic activity [ 42 ]. Highly pure and folded rAmb a 6 was obtained exhibiting the expected molecular mass according to its amino acid sequence.…”

Section: Discussionmentioning

confidence: 99%

“…Cells were pre-incubated overnight with patient serum heat-inactivated at 55 °C for 25 min [ 57 ] diluted 1:10 in MEM supplemented with 10% heat-inactivated fetal bovine serum (HI FBS), 100 U/mL penicillin–streptomycin, 0.2 mM L-glutamine, 0.2 mg/mL hygromycin B, and 0.2 mg/mL geneticin (Thermo Fisher Scientific). Six different allergen concentrations (1000, 100, 10, 1, 0.1, and 0.01 ng/mL) from rAmb a 6; Par j 2, kindly provided by Prof. Rudolf Valenta [ 42 ]; and natural Amb a 1.01 (nAmb a 1.01) as a benchmark kindly provided by Dr. Frank Stolz were used to test allergenic activity. The assay was performed with minor modifications from the previously described assay [ 58 ], namely exposure to the allergen for 1 h at 37 °C, 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

Non-Specific Lipid Transfer Protein Amb a 6 Is a Source-Specific Important Allergenic Molecule in Ragweed Pollen

Grijincu,

Tănasie,

Zbîrcea

et al. 2024

IJMS

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Pollen from common ragweed is an important allergen source worldwide and especially in western and southern Romania. More than 100 million patients suffer from symptoms of respiratory allergy (e.g., rhinitis, asthma) to ragweed pollen. Among the eleven characterized allergens, Amb a 6 is a non-specific lipid transfer protein (nsLTP). nsLTPs are structurally stable proteins in pollen and food from different unrelated plants capable of inducing severe reactions. The goal of this study was to produce Amb a 6 as a recombinant and structurally folded protein (rAmb a 6) and to characterize its physicochemical and immunological features. rAmb a 6 was expressed in Spodoptera frugiperda Sf9 cells as a secreted protein and characterized by mass spectrometry and circular dichroism (CD) spectroscopy regarding molecular mass and fold, respectively. The IgE-binding frequency towards the purified protein was evaluated using sera from 150 clinically well-characterized ragweed-allergic patients. The allergenic activities of rAmb a 6 and the nsLTP from the weed Parietaria judaica (Par j 2) were evaluated in basophil activation assays. rAmb a 6-specific IgE reactivity was associated with clinical features. Pure rAmb a 6 was obtained by insect cell expression. Its deduced molecular weight corresponded to that determined by mass spectrometry (i.e., 10,963 Da). rAmb a 6 formed oligomers as determined by SDS-PAGE under non-reducing conditions. According to multiple sequence comparisons, Amb a 6 was a distinct nsLTP with less than 40% sequence identity to currently known plant nsLTP allergens, except for nsLTP from Helianthus (i.e., 52%). rAmb a 6 is an important ragweed allergen recognized by 30% of ragweed pollen allergic patients. For certain patients, rAmb a 6-specific IgE levels were higher than those specific for the major ragweed allergen Amb a 1 and analysis also showed a higher allergenic activity in the basophil activation test. rAmb a 6-positive patients suffered mainly from respiratory symptoms. The assumption that Amb a 6 is a source-specific ragweed allergen is supported by the finding that none of the patients showing rAmb a 6-induced basophil activation reacted with Par j 2 and only one rAmb a 6-sensitized patient had a history of plant food allergy. Immunization of rabbits with rAmb a 6 induced IgG antibodies which strongly inhibited IgE binding to rAmb a 6. Our results demonstrate that Amb a 6 is an important source-specific ragweed pollen allergen that should be considered for diagnosis and allergen-specific immunotherapy of ragweed pollen allergy.

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“…93,[239][240][241] For instance, basophil activation induced by mix of increasing concentrations of the four major timothy grass pollen allergens (rPhl p 1, 2, 5, and 6) was reduced during recombinant B cell epitope-based vaccine (BM32) immunotherapy of patients with grass pollen allergy. 240 Furthermore, rPhl p 5 dependent basophil activation inhibition with SCIT sera demonstrated that immunotherapy-induced allergen-specific IgG antibodies are not long-lasting after treatment discontinuation. 242 In weed pollen allergy, Parietaria judaica represents one of the main sources of allergens in the Mediterranean area with Par j 1 and Par j 2 as major allergens.…”

Section: Grass and Weed Pollen Allergensmentioning

confidence: 99%

“…221 BAT is important for the characterization of novel recombinant, hypoallergenic, peptide-based vaccines for grass pollen allergy and has become a major tool for evaluating change in allergenicity when basophils from patients allergic to grass pollen are tested with novel vaccines peptides and/or carriers. 93,[239][240][241] For instance, basophil activation induced by mix of increasing concentrations of the four major timothy grass pollen allergens (rPhl p 1, 2, 5, and 6) was reduced during recombinant B cell epitope-based vaccine (BM32) immunotherapy of patients with grass pollen allergy. 240 Furthermore, rPhl p 5 dependent basophil activation inhibition with SCIT sera demonstrated that immunotherapy-induced allergen-specific IgG antibodies are not long-lasting after treatment discontinuation.…”

Section: Grass and Weed Pollen Allergensmentioning

confidence: 99%

EAACIMolecular Allergology User's Guide 2.0

Dramburg

Hilger

Santos

et al. 2023

Pediatric Allergy Immunology

101

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Since the discovery of immunoglobulin E (IgE) as a mediator of allergic diseases in 1967, our knowledge about the immunological mechanisms of IgE‐mediated allergies has remarkably increased. In addition to understanding the immune response and clinical symptoms, allergy diagnosis and management depend strongly on the precise identification of the elicitors of the IgE‐mediated allergic reaction. In the past four decades, innovations in bioscience and technology have facilitated the identification and production of well‐defined, highly pure molecules for component‐resolved diagnosis (CRD), allowing a personalized diagnosis and management of the allergic disease for individual patients. The first edition of the “EAACI Molecular Allergology User's Guide” (MAUG) in 2016 rapidly became a key reference for clinicians, scientists, and interested readers with a background in allergology, immunology, biology, and medicine. Nevertheless, the field of molecular allergology is moving fast, and after 6 years, a new EAACI Taskforce was established to provide an updated document. The Molecular Allergology User's Guide 2.0 summarizes state‐of‐the‐art information on allergen molecules, their clinical relevance, and their application in diagnostic algorithms for clinical practice. It is designed for both, clinicians and scientists, guiding health care professionals through the overwhelming list of different allergen molecules available for testing. Further, it provides diagnostic algorithms on the clinical relevance of allergenic molecules and gives an overview of their biology, the basic mechanisms of test formats, and the application of tests to measure allergen exposure.

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

Cited by 6 publications

References 49 publications

Preventive Allergen-Specific Vaccination Against Allergy: Mission Possible?

Preventive Allergen-Specific Vaccination Against Allergy: Mission Possible?

Non-Specific Lipid Transfer Protein Amb a 6 Is a Source-Specific Important Allergenic Molecule in Ragweed Pollen

EAACIMolecular Allergology User's Guide 2.0

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