Cloning, isolation, and IgE-binding properties of Helix aspersa (brown garden snail) tropomyosin

Int Arch Allergy Immunol. 2002 Jun;128(2):90-6. doi: 10.1159/000059398.

Abstract

Background: Gastropod consumption is quite frequent in the Mediterranean countries and cross-reactivities with crustaceans have been described, but the mechanism of this allergenic cross-reactivity has not been studied in detail. This study aimed to produce recombinant Helix aspersa (brown garden snail) tropomyosin and investigate its implication for cross-reactivity among invertebrates.

Methods: A tropomyosin-specific cDNA encoding H. aspersa tropomyosin was synthetized, and recombinant allergen was overexpressed in Escherichia coli as nonfusion protein. IgE-binding reactivity was studied by immunoblotting and immunoblot inhibition experiments with sera from snail-allergic patients.

Results: Cloned brown garden snail tropomyosin shares high homology with other edible mollusk tropomyosins (84-69% identity) as well as with those from arthropods (65-62%), and less homology with vertebrate ones (56% identity). Tropomyosin reacted with 18% of the sera from patients with snail allergy. Inhibition experiments, using natural and recombinant tropomyosins, showed different degrees of cross-reactivity between invertebrate tropomyosins. Sera from snail-allergic subjects recognized tropomyosins in both mollusks and crustacean extracts.

Conclusions: Tropomyosin represents a minor allergen in snail extracts, but it is clearly involved in invertebrate cross-reactivity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Western
  • Cloning, Molecular
  • Cross Reactions
  • DNA, Complementary / chemistry
  • DNA, Complementary / genetics
  • Escherichia coli / genetics
  • Food Hypersensitivity / blood
  • Food Hypersensitivity / immunology*
  • Helix, Snails / genetics
  • Helix, Snails / immunology*
  • Humans
  • Immunoglobulin E / immunology*
  • Immunoglobulin E / metabolism
  • Molecular Sequence Data
  • Mollusca
  • Muscles / chemistry
  • Muscles / immunology
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Homology, Amino Acid
  • Tissue Extracts / immunology
  • Tropomyosin / genetics
  • Tropomyosin / immunology*
  • Tropomyosin / isolation & purification

Substances

  • DNA, Complementary
  • RNA, Messenger
  • Recombinant Proteins
  • Tissue Extracts
  • Tropomyosin
  • Immunoglobulin E