Andrea Wangorsch1, Anuja Kulkarni1,2, Annette Jamin1, Jelena Spiric1, Julia Bräcker3, Jens Brockmeyer3, Vera Mahler1, Natalia Blanca-López4, Marta Ferrer5, Miguel Blanca4,6, Maria Torres6, Paqui Gomez6, Joan Bartra7,8, Alba García-Moral7, María J Goikoetxea5, Stefan Vieths1, Masako Toda1,9, Gianni Zoccatelli10, Stephan Scheurer1. 1. Division Allergology and Section Molecular Allergology, Paul-Ehrlich-Institut, Paul-Ehrlich-Str. 2, 63225, Langen, Germany. 2. Amity University, Mumbai, India. 3. Analytical Food Chemistry, University of Stuttgart, Allmandring 5B, 70569, Stuttgart, Germany. 4. Allergy Service, Hospital Infanta Leonor, Gran Via del Este 80, Madrid, 28031, Spain. 5. Department of Allergy, IdiSNA (Instituto de Investigación Sanitaria de Navarra), Clinica Universidad de Navarra, Pio XII, Pamplona, 3631008, Spain. 6. Jefe de Servicio de Alergología, Hospital Civil, Plaza del Hospital Civil s/n, Pabellon 5, sotano, Málaga, 29009, Spain. 7. Allergy Unit, Pneumology Department, Clinic Hospital, Sant Antoni Maria Claret, 167, Barcelona, Catalunya, 08025, Spain. 8. Institut d'Investigacions Biomediques August Pi i Sunyer (IDIBAPS), University of Barcelona, Carrer del Rosselló, 149, Barcelona, 08036, Spain. 9. Laboratory of Food and Biomolecular Science, Graduate School of Agricultural Science, Tohoku University, Aramaki 468-1, Aoba-ku, Sendai-city, 980-8572, Japan. 10. Department of Biotechnology, University of Verona, Strada le Grazie 15, Verona, 37134, Italy.
Abstract
SCOPE: Chickpea (Cicer arietinum) allergy has frequently been reported particularly in Spain and India. Nevertheless, chickpea allergens are poorly characterized. The authors aim to identify and characterize potential allergens from chickpea. METHODS AND RESULTS: Candidate proteins are selected by an in silico approach or immunoglobuline E (IgE)-testing. Potential allergens are prepared as recombinant or natural proteins and characterized for structural integrity by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), circular dichroism (CD)-spectroscopy, and mass spectrometry (MS) analysis. IgE-sensitization pattern of Spanish chickpea allergic and German peanut and birch pollen sensitized patients are investigated using chickpea extracts and purified proteins. Chickpea allergic patients show individual and heterogeneous IgE-sensitization profiles with extracts from raw and boiled chickpeas. Chickpea proteins pathogenesis related protein family 10 (PR-10), a late embryogenesis abundant protein (LEA/DC-8), and a vicilin-containing fraction, but not 2S albumin, shows IgE reactivity with sera from chickpea, birch pollen, and peanut sensitized patients. Remarkably, allergenic vicilin, DC-8, and PR-10 are detected in the extract of boiled chickpeas. CONCLUSION: Several IgE-reactive chickpea allergens are identified. For the first time a yet not classified DC-8 protein is characterized as minor allergen (Cic a 1). Finally, the data suggest a potential risk for peanut allergic patients by IgE cross-reactivity with homologous chickpea proteins.
SCOPE: Chickpea (Cicer arietinum) allergy has frequently been reported particularly in Spain and India. Nevertheless, chickpea allergens are poorly characterized. The authors aim to identify and characterize potential allergens from chickpea. METHODS AND RESULTS: Candidate proteins are selected by an in silico approach or immunoglobuline E (IgE)-testing. Potential allergens are prepared as recombinant or natural proteins and characterized for structural integrity by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), circular dichroism (CD)-spectroscopy, and mass spectrometry (MS) analysis. IgE-sensitization pattern of Spanish chickpea allergic and German peanut and birch pollen sensitized patients are investigated using chickpea extracts and purified proteins. Chickpea allergicpatients show individual and heterogeneous IgE-sensitization profiles with extracts from raw and boiled chickpeas. Chickpea proteins pathogenesis related protein family 10 (PR-10), a late embryogenesis abundant protein (LEA/DC-8), and a vicilin-containing fraction, but not 2S albumin, shows IgE reactivity with sera from chickpea, birch pollen, and peanut sensitized patients. Remarkably, allergenic vicilin, DC-8, and PR-10 are detected in the extract of boiled chickpeas. CONCLUSION: Several IgE-reactive chickpea allergens are identified. For the first time a yet not classified DC-8 protein is characterized as minor allergen (Cic a 1). Finally, the data suggest a potential risk for peanutallergicpatients by IgE cross-reactivity with homologous chickpea proteins.