Myosin light chain is a novel shrimp allergen, Lit v 3

• Published in: Journal of allergy and clinical immunology Vol. 122; no. 4; pp. 795 - 802
• Main Authors: Ayuso, Rosalía, Grishina, Galina, Bardina, Ludmilla, Carrillo, Teresa, Blanco, Carlos, Ibáñez, María Dolores, Sampson, Hugh A., Beyer, Kirsten
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01.10.2008; Elsevier; Elsevier Limited
• Subjects: Adolescent; Adult; allergen; Allergens - genetics; Allergens - immunology; Allergies; Allergy and Immunology; Animals; Biological and medical sciences; Child; crustacean; Crustaceans; Female; Food allergies; Food Hypersensitivity - blood; Food Hypersensitivity - immunology; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Gene Library; Humans; Immunoglobulin E - blood; Immunoglobulin E - immunology; Lit v 3; Litopenaeus vannamei; Male; Marine; Medical sciences; Myosin light chain; Myosin Light Chains - genetics; Myosin Light Chains - immunology; Penaeidae - genetics; Penaeidae - immunology; recombinant; Recombinant Proteins - genetics; Recombinant Proteins - immunology; Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis; Shellfish; Shellfish - adverse effects; shrimp; United States > US; I, Pacific; allergen; shrimp; crustacean; MLC; MALDI; MS; recombinant; Lit v 3; 2-D; Myosin light chain; Two-dimensional; Matrix-assisted laser desorption/ionization; Mass spectrometry; Allergy; Immunopathology; Contractile protein; Light peptide chain; Macrura; Crustacea; Immunology; Arthropoda; Myosin; Decapoda; Recombinant protein; Invertebrata; Shrimp; Allergen
• ISSN: 0091-6749; 1097-6825; 1097-6825
• DOI: 10.1016/j.jaci.2008.07.023

Shellfish allergy is a prevalent, long-lasting disorder usually persisting throughout life. Few options are available for treatment, and avoidance is the only therapy recommended. We sought to identify relevant crustacean allergens for use as diagnostic and safe immunotherapeutic agents for subjects with shellfish allergy. Thirty-eight patients were selected with immediate allergic reactions to shrimp and increased shrimp-specific serum IgE levels. One-dimensional and 2-dimensional electrophoresis of shrimp extracts were followed by IgE immunoblotting. Protein identification was done with matrix-assisted laser desorption/ionization–mass spectrometry and Edman sequencing. A cDNA library was generated from white pacific shrimp (Litopenaeus vannamei) and screened with primers designed on the basis of internal sequences obtained from 2-dimensional tryptic digests. Full-length cDNA clones were isolated from the library and sequenced. Recombinant protein was expressed and tested with sera from patients with shrimp allergy. Immunoblotting demonstrated IgE binding to a 20-kDa shrimp protein by 21 (55%) of 38 sera. Tryptic digestion of the protein followed by matrix-assisted laser desorption/ionization–mass spectrometric analysis and Edman sequencing identified it as a myosin light chain (MLC). Screening of the shrimp cDNA library resulted in isolation of a novel protein cDNA. Open reading frame translation provided the amino acid sequence of a new allergenic shrimp protein with high similarity to Bla g 8 (cockroach MLC). Recombinant protein was recognized by 17 patients, confirming the allergenicity of shrimp MLC. We have identified and cloned a new major shrimp allergen, Lit v 3.0101, an MLC protein.