title: Generation of a comprehensive panel of crustacean allergens from the North Sea Shrimp Crangon crangon.
abstract: BACKGROUND
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Paul-Ehrlich-Institut, Division of Allergology, Paul-Ehrlich-Str. 51-59, D-63225 Langen, Germany.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Bauermeister
authors.forename: Andrea
authors.initials: A
authors.lastname: Wangorsch
authors.forename: Lorenza Perono
authors.initials: LP
authors.lastname: Garoffo
authors.forename: Andreas
authors.initials: A
authors.lastname: Reuter
authors.forename: Amedeo
authors.initials: A
authors.lastname: Conti
authors.forename: Steve L
authors.initials: SL
authors.lastname: Taylor
authors.forename: Jonas
authors.initials: J
authors.lastname: Lidholm
authors.forename: Asa Marknell
authors.initials: AM
authors.lastname: Dewitt
authors.forename: Ernesto
authors.initials: E
authors.lastname: Enrique
authors.forename: Stefan
authors.initials: S
authors.lastname: Vieths
authors.forename: Thomas
authors.initials: T
authors.lastname: Holzhauser
authors.forename: Barbara
authors.initials: B
authors.lastname: Ballmer-Weber
authors.forename: Gerald
authors.initials: G
authors.lastname: Reese
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000485:Allergens / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002648:Child; D033501:Crangonidae / Q000737:chemistry / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005512:Food Hypersensitivity / Q000175:diagnosis* / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D055815:Young Adult
chemical_list: D000485:Allergens; D011994:Recombinant Proteins; D007073:Immunoglobulin E