Generation of a comprehensive panel of crustacean allergens from the North Sea Shrimp Crangon crangon

• Published in: Molecular immunology Vol. 48; no. 15-16; pp. 1983 - 1992
• Main Authors: Bauermeister, Kerstin, Wangorsch, Andrea, Garoffo, Lorenza Perono, Reuter, Andreas, Conti, Amedeo, Taylor, Steve L., Lidholm, Jonas, DeWitt, Åsa Marknell, Enrique, Ernesto, Vieths, Stefan, Holzhauser, Thomas, Ballmer-Weber, Barbara, Reese, Gerald
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.09.2011
• Subjects: Adolescent; Adult; Aged; allergens; Allergens - chemistry; Allergens - immunology; Allergens - isolation & purification; Amino Acid Sequence; anaphylaxis; Animals; Arginine kinase; biomarkers; Blotting, Western; Child; complementary DNA; Crangon crangon; Crangonidae - chemistry; Crangonidae - immunology; Crustacean allergy; diagnostic sensitivity; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Europe; EuroPrevall; Female; food allergies; Food Hypersensitivity - diagnosis; Food Hypersensitivity - immunology; Humans; immunoglobulin E; Immunoglobulin E - blood; Male; Marine; Mass Spectrometry; messenger RNA; Middle Aged; Myosin light chain; myosin light chains; polyacrylamide gel electrophoresis; recombinant proteins; Recombinant Proteins - immunology; Sarcoplasmic Ca-binding protein; Sensitivity and Specificity; sequence homology; shrimp; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; triose-phosphate isomerase; Triosephosphate isomerase; Tropomyosin; tropomyosins; Troponin C; vaccination; Young Adult; Europe; ANE, North Sea; ANE, Europe; FPLC; SCP; Crustacean allergy; IMAC; MALDI-TOF; Crangon crangon; MLC; Tropomyosin; MS; Triosephosphate isomerase; TnC; AK; Myosin light chain; EuroPrevall; LC; Arginine kinase; TIM; TM; Troponin C; Sarcoplasmic Ca-binding protein
• ISSN: 0161-5890; 1872-9142; 1872-9142
• DOI: 10.1016/j.molimm.2011.06.216

► Published data on crustacean allergens are incomplete. ► We identify, clone, and confirm six shrimp allergens, three of which are novel. ► A combined in vitro diagnostic sensitivity is similar to established shrimp extract. ► This allergen panel forms a basis for future efforts on potential biomarkers. ► Allergen specific patient-tailored immunotherapeutic strategies might be approached. Published data on crustacean allergens are incomplete. The identification of tropomyosin (TM), arginine kinase (AK), sarcoplasmic Ca-binding protein (SCP) and myosin light chain (MLC) as shrimp allergens are all important contributions but additional allergens are required for the development of a complete set of reagents for component resolved diagnosis and the exploration of novel vaccination strategies. The North Sea shrimp (Crangon crangon), which is frequently consumed in Europe, served as a model organism in this study. TM and AK were directly cloned from mRNA based on sequence homology and produced as recombinant proteins. Additional IgE-reactive proteins were isolated by preparative SDS-PAGE and identified by mass spectrometry and corresponding cDNAs were cloned and expressed in E. coli. The relevance of the 6 cloned crustacean allergens was confirmed with sera of 31 shrimp-allergic subjects, 12 of which had a positive double-blind, placebo-controlled food challenge (DBPCFC) to shrimp and 19 a convincing history of food allergy to shrimp, including 5 cases of anaphylaxis. Quantitative IgE measurements were performed by ImmunoCAP. Six recombinant crustacean proteins: TM, AK, SCP, a novel MLC, troponin C (TnC), and triosephosphate isomerase (TIM) bound IgE in ImmunoCAP analysis. Specific IgE to at least one of these single shrimp allergens was detected in 90% of the study population, thus the in vitro diagnostic sensitivity was comparable to that of shrimp extract (97%). In 75% of the subjects, the combined technical sensitivity was similar to or greater with single shrimp allergens than with natural shrimp extract. We identified six IgE-binding proteins from C. crangon, three of which have not before been described as allergens in crustaceans. This extensive panel of shrimp allergens forms a valuable asset for future efforts towards the identification of clinically relevant biomarkers and as a basis to approach patient-tailored immunotherapeutic strategies.