Generation of a comprehensive panel of crustacean allergens from the North Sea Shrimp Crangon crangon.
PMID 21784530 · Molecular immunology · 2011 · DOI 10.1016/j.molimm.2011.06.216
Authors: Kerstin Bauermeister; Andrea Wangorsch; Lorenza Perono Garoffo; Andreas Reuter; Amedeo Conti; Steve L Taylor; Jonas Lidholm; Asa Marknell Dewitt; Ernesto Enrique; Stefan Vieths; Thomas Holzhauser; Barbara Ballmer-Weber; Gerald Reese
Abstract
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.
Adolescent; Adult; Aged; Allergens; Amino Acid Sequence; Animals; Blotting, Western; Child; Crangonidae; Electrophoresis, Polyacrylamide Gel; Female; Food Hypersensitivity; Humans; Immunoglobulin E; Male; Mass Spectrometry; Middle Aged; Recombinant Proteins; Sensitivity and Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Young Adult
Cite
Kerstin Bauermeister; Andrea Wangorsch; Lorenza Perono Garoffo; Andreas Reuter; Amedeo Conti; Steve L Taylor; Jonas Lidholm; Asa Marknell Dewitt; Ernesto Enrique; Stefan Vieths; Thomas Holzhauser; Barbara Ballmer-Weber; Gerald Reese. Generation of a comprehensive panel of crustacean allergens from the North Sea Shrimp Crangon crangon. Molecular immunology (2011). PMID 21784530. DOI 10.1016/j.molimm.2011.06.216.