TY - JOUR
T1 - Art v 1 IgE epitopes of patients and humanized mice are conformational
AU - Zabel, Maja
AU - Weber, Milena
AU - Kratzer, Bernhard
AU - Köhler, Cordula
AU - Jahn-Schmid, Beatrice
AU - Gadermaier, Gabriele
AU - Gattinger, Pia
AU - Bidovec-Stojkovič, Urška
AU - Korošec, Peter
AU - Smole, Ursula
AU - Wurzinger, Gert
AU - Chen, Kuan-Wei
AU - Panaitescu, Carmen Bunu
AU - Klimek, Ludger
AU - Pablos, Isabel
AU - Niespodziana, Katarzyna
AU - Neunkirchner, Alina
AU - Keller, Walter
AU - Valenta, Rudolf
AU - Pickl, Winfried F
N1 - Funding Information:
This work was supported by the Austrian Science Foundation (FWF; grant nos. DK-W1248 and SFB F4609 ), by the DANUBE-ARC grant of the country of Lower Austria, and partially by a grant from WORG Pharmaceuticals.
Funding Information:
Disclosure of potential conflict of interest: W. F. Pickl reports grants from the Austrian Science Fund and the Country of Lower Austria, during the conduct of the study, as well as personal fees from Novartis (Vienna, Austria) outside the submitted work. R. Valenta reports grants and personal fees from Viravaxx (Vienna, Austria) as well as grants from HVD Biotech (Vienna, Austria) and WORG Pharmaceuticals (Hangzhou, China), all outside the submitted work. In addition, W. F. Pickl, R. Valenta, and M. Zabel have a patent regarding IgE binding sites pending. G. Gadermaier reports personal fees from Bencard (Vienna, Austria), outside the submitted work. G. Wurzinger reports personal fees from Novartis (Base, Switzerland), Chiesi (Parma, Italy), ThermoFisherScientific (Waltham, Mass), and Allergopharma (Reinbek, Germany), outside the submitted work. L. Klimek reports grants and personal fees from Allergopharma (Reinbek, Germany), MEDA/Mylan (Solna, Sweden), LETI Pharma (Barcelona, Spain), Stallergenes (London, UK), Sanofi (Paris, France), and GlaxoSmithKline (Brentford, UK); grants from Quintiles (Durham, NC), ASIT Biotech (Liège, Belgium), Lofarma (Milano, Italy), AstraZeneca (Cambridge, UK), and Inmunotek (Madrid, Spain); personal fees from HAL Allergie (Leiden, Netherlands), ALK Abelló (Hørsholm, Denmark), Allergy Therapeut. (Worthing, UK), Cassella (Frankfurt, Germany), med and Novartis (Basel, Switzerland), as well as several memberships (AeDA, DGHNO, Deutsche Akademie für Allergologie und klinische Immunologie, HNO-BV, GPA, and EAACI), all outside the submitted work. W. Keller reports grants from the Austrian Science Fund (FWF), during the conduct of the study. The rest of the authors declare that they have no additional financial interests.
Funding Information:
This work was supported by the Austrian Science Foundation (FWF; grant nos. DK-W1248 and SFB F4609), by the DANUBE-ARC grant of the country of Lower Austria, and partially by a grant from WORG Pharmaceuticals. Disclosure of potential conflict of interest: W. F. Pickl reports grants from the Austrian Science Fund and the Country of Lower Austria, during the conduct of the study, as well as personal fees from Novartis (Vienna, Austria) outside the submitted work. R. Valenta reports grants and personal fees from Viravaxx (Vienna, Austria) as well as grants from HVD Biotech (Vienna, Austria) and WORG Pharmaceuticals (Hangzhou, China), all outside the submitted work. In addition, W. F. Pickl, R. Valenta, and M. Zabel have a patent regarding IgE binding sites pending. G. Gadermaier reports personal fees from Bencard (Vienna, Austria), outside the submitted work. G. Wurzinger reports personal fees from Novartis (Base, Switzerland), Chiesi (Parma, Italy), ThermoFisherScientific (Waltham, Mass), and Allergopharma (Reinbek, Germany), outside the submitted work. L. Klimek reports grants and personal fees from Allergopharma (Reinbek, Germany), MEDA/Mylan (Solna, Sweden), LETI Pharma (Barcelona, Spain), Stallergenes (London, UK), Sanofi (Paris, France), and GlaxoSmithKline (Brentford, UK); grants from Quintiles (Durham, NC), ASIT Biotech (Liège, Belgium), Lofarma (Milano, Italy), AstraZeneca (Cambridge, UK), and Inmunotek (Madrid, Spain); personal fees from HAL Allergie (Leiden, Netherlands), ALK Abelló (Hørsholm, Denmark), Allergy Therapeut. (Worthing, UK), Cassella (Frankfurt, Germany), med and Novartis (Basel, Switzerland), as well as several memberships (AeDA, DGHNO, Deutsche Akademie für Allergologie und klinische Immunologie, HNO-BV, GPA, and EAACI), all outside the submitted work. W. Keller reports grants from the Austrian Science Fund (FWF), during the conduct of the study. The rest of the authors declare that they have no additional financial interests.
Publisher Copyright:
© 2022 The Authors
PY - 2022/10
Y1 - 2022/10
N2 - BACKGROUND: Worldwide, pollen of the weed mugwort (Artemisiavulgaris) is a major cause of severe respiratory allergy, with its major allergen, Art v 1, being the key pathogenic molecule for millions of patients. Humanized mice transgenic for a human T-cell receptor specific for the major Art v 1 T-cell epitope and the corresponding HLA have been made.OBJECTIVE: We sought to characterize IgE epitopes of Art v 1-sensitized patients and humanized mice for molecular immunotherapy of mugwort allergy.METHODS: Four overlapping peptides incorporating surface-exposed amino acids representing the full-length Art v 1 sequence were synthesized and used to search for IgE reactivity to sequential epitopes. For indirect mapping, peptide-specific rabbit antibodies were raised to block IgE against surface-exposed epitopes on folded Art v 1. IgE reactivity and basophil activation studies were performed in clinically defined mugwort-allergic patients. Secondary structure of recombinant (r) Art v 1 and peptides was determined by circular dichroism spectroscopy.RESULTS: Mugwort-allergic patients and humanized mice sensitized by allergen inhalation showed IgE reactivity and/or basophil activation mainly to folded, complete Art v 1 but not to unfolded, sequential peptide epitopes. Blocking of allergic patients' IgE with peptide-specific rabbit antisera identified a hitherto unknown major conformational IgE binding site in the C-terminal Art v 1 domain.CONCLUSIONS: Identification of the new major conformational IgE binding site on Art v 1, which can be blocked with IgG raised against non-IgE reactive Art v 1 peptides, is an important basis for the development of a hypoallergenic peptide vaccine for mugwort allergy.
AB - BACKGROUND: Worldwide, pollen of the weed mugwort (Artemisiavulgaris) is a major cause of severe respiratory allergy, with its major allergen, Art v 1, being the key pathogenic molecule for millions of patients. Humanized mice transgenic for a human T-cell receptor specific for the major Art v 1 T-cell epitope and the corresponding HLA have been made.OBJECTIVE: We sought to characterize IgE epitopes of Art v 1-sensitized patients and humanized mice for molecular immunotherapy of mugwort allergy.METHODS: Four overlapping peptides incorporating surface-exposed amino acids representing the full-length Art v 1 sequence were synthesized and used to search for IgE reactivity to sequential epitopes. For indirect mapping, peptide-specific rabbit antibodies were raised to block IgE against surface-exposed epitopes on folded Art v 1. IgE reactivity and basophil activation studies were performed in clinically defined mugwort-allergic patients. Secondary structure of recombinant (r) Art v 1 and peptides was determined by circular dichroism spectroscopy.RESULTS: Mugwort-allergic patients and humanized mice sensitized by allergen inhalation showed IgE reactivity and/or basophil activation mainly to folded, complete Art v 1 but not to unfolded, sequential peptide epitopes. Blocking of allergic patients' IgE with peptide-specific rabbit antisera identified a hitherto unknown major conformational IgE binding site in the C-terminal Art v 1 domain.CONCLUSIONS: Identification of the new major conformational IgE binding site on Art v 1, which can be blocked with IgG raised against non-IgE reactive Art v 1 peptides, is an important basis for the development of a hypoallergenic peptide vaccine for mugwort allergy.
KW - Allergy
KW - Art v 1
KW - IgE epitope
KW - allergen
KW - allergen-specific immunotherapy
KW - epitope mapping
KW - mugwort pollen allergy
UR - http://www.scopus.com/inward/record.url?scp=85132708710&partnerID=8YFLogxK
U2 - 10.1016/j.jaci.2022.04.031
DO - 10.1016/j.jaci.2022.04.031
M3 - Journal article
C2 - 35738928
SN - 0091-6749
VL - 150
SP - 920
EP - 930
JO - Journal of Allergy and Clinical Immunology
JF - Journal of Allergy and Clinical Immunology
IS - 4
ER -