Rosalía Ayuso1, Galina Grishina2, Ludmilla Bardina2, Teresa Carrillo3, Carlos Blanco4, María Dolores Ibáñez5, Hugh A Sampson2, Kirsten Beyer6. 1. Division of Allergy and Immunology and the Jaffe Food Allergy Research Institute, Mount Sinai School of Medicine, New York, NY. Electronic address: rosalia.ayuso@mssm.edu. 2. Division of Allergy and Immunology and the Jaffe Food Allergy Research Institute, Mount Sinai School of Medicine, New York, NY. 3. Section of Allergy and Respiratory Diseases, Hospital Universitario Dr. Negrín, Las Palmas de Gran Canaria, Spain. 4. Section of Allergy, Hospital Universitario de la Princesa, Madrid, Spain. 5. Pediatric Allergy, Hospital del Niño Jesús, Madrid, Spain. 6. Department of Pediatric Pneumology and Immunology, University Children's Hospital Charité, Berlin, Germany.
Abstract
BACKGROUND: 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. OBJECTIVE: We sought to identify relevant crustacean allergens for use as diagnostic and safe immunotherapeutic agents for subjects with shellfish allergy. METHODS: 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. RESULTS: 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. CONCLUSIONS: We have identified and cloned a new major shrimp allergen, Lit v 3.0101, an MLC protein.
BACKGROUND: 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. OBJECTIVE: We sought to identify relevant crustacean allergens for use as diagnostic and safe immunotherapeutic agents for subjects with shellfish allergy. METHODS: 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. RESULTS: 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. CONCLUSIONS: We have identified and cloned a new major shrimp allergen, Lit v 3.0101, an MLC protein.
Authors: L Karla Arruda; Michelle C R Barbosa; Ana Beatriz R Santos; Adriana S Moreno; Martin D Chapman; Anna Pomés Journal: Curr Allergy Asthma Rep Date: 2014-04 Impact factor: 4.806
Authors: Nicki Y H Leung; Christine Y Y Wai; ShangAn Shu; Jinjun Wang; Thomas P Kenny; Ka Hou Chu; Patrick S C Leung Journal: Clin Rev Allergy Immunol Date: 2014-06 Impact factor: 8.667