The Journal of Allergy and Clinical Immunology
Volume 122, Issue 4 , Pages 795-802, October 2008

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

  • Rosalía Ayuso, MD, PhD

      Affiliations

    • Division of Allergy and Immunology and the Jaffe Food Allergy Research Institute, Mount Sinai School of Medicine, New York, NY
    • Corresponding Author InformationReprint requests: Rosalía Ayuso, MD, PhD, Mount Sinai School of Medicine, One Gustave L. Levy Place, Box 1198, New York, NY 10029.
    • ,
  • Galina Grishina, MS

      Affiliations

    • Division of Allergy and Immunology and the Jaffe Food Allergy Research Institute, Mount Sinai School of Medicine, New York, NY
    • ,
  • Ludmilla Bardina, MS

      Affiliations

    • Division of Allergy and Immunology and the Jaffe Food Allergy Research Institute, Mount Sinai School of Medicine, New York, NY
    • ,
  • Teresa Carrillo, MD, PhD

      Affiliations

    • Section of Allergy and Respiratory Diseases, Hospital Universitario Dr. Negrín, Las Palmas de Gran Canaria, Spain
    • ,
  • Carlos Blanco, MD, PhD

      Affiliations

    • Section of Allergy, Hospital Universitario de la Princesa, Madrid, Spain
    • ,
  • María Dolores Ibáñez, MD, PhD

      Affiliations

    • Pediatric Allergy, Hospital del Niño Jesús, Madrid, Spain
    • ,
  • Hugh A. Sampson, MD

      Affiliations

    • Division of Allergy and Immunology and the Jaffe Food Allergy Research Institute, Mount Sinai School of Medicine, New York, NY
    • ,
  • Kirsten Beyer, MD

      Affiliations

    • Department of Pediatric Pneumology and Immunology, University Children's Hospital Charité, Berlin, Germany

Received 22 April 2008; received in revised form 16 July 2008; accepted 18 July 2008. published online 29 August 2008.

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.

Key words: Myosin light chain, allergen, crustacean, shrimp, recombinant, Lit v 3

Abbreviations used: 2-D, Two-dimensional, MALDI, Matrix-assisted laser desorption/ionization, MLC, Myosin light chain, MS, Mass spectrometry

 

 Disclosure of potential conflict of interest: R. Ayuso has received research support from the Food Allergy Initiative. H. A. Sampson has consulting arrangements with Allertein Therapeutics, Schering, and the Food Allergy Initiative and has received research support from the National Institute of Allergy and Infectious Diseases, the National Institutes of Health, and the Food Allergy Initiative. K. Beyer has received research support from the European Union, the Food Allergy and Anaphylaxis Network, Phadia, and the Paul Ehrlich Institute. The rest of the authors have declared that they have no conflict of interest.

PII: S0091-6749(08)01352-3

doi:10.1016/j.jaci.2008.07.023

The Journal of Allergy and Clinical Immunology
Volume 122, Issue 4 , Pages 795-802, October 2008

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