| Autor: |
R. Iglesias, E. García-Bodas, A. M. Anadón, Esperanza Rodríguez, Teresa Gárate, F. Romarís, Florencio M. Ubeira |
| Rok vydání: |
2008 |
| Předmět: |
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| Zdroj: |
Allergy. 63:219-225 |
| ISSN: |
0105-4538 |
| DOI: |
10.1111/j.1398-9995.2007.01564.x |
| Popis: |
Background: Anisakis simplex allergens may cause severe allergic reactions in infected patients. Human anisakiasis can be specifically diagnosed by detection of immunoglobulin E (IgE) antibodies against O-deglycosylated nAni s 7 allergen captured by monoclonal antibody (mAb) UA3 (UA3-ELISA), although the nature of this important allergen is unknown. The aim of this study was to clone and characterize the Ani s 7 major allergen, and to obtain a recombinant fragment suitable for serodiagnosis. Methods: An Anisakis cDNA library was screened with mAb UA3 and a cDNA clone (rAni s 7) encoding a 1096-amino acid fragment of Ani s 7 (GenBank: EF158010) was identified. Bioinformatic tools and immunological and biochemical techniques were used to characterize the allergen obtained. Results: The rAni s 7 fragment comprised 19 repeats of a novel CX17−25CX9−22CX8CX6 tandem repeat motif not seen in any previously reported protein sequence. An internal 435Met–713Arg fragment of the rAni s 7 (t-Ani s 7) was expressed in Escherichia coli and evaluated for serodiagnostic utility. Indirect enzyme-linked immunosorbent assay (ELISA) with t-Ani s 7 identified as positive the same 60 sera as UA3-ELISA. The sequence MCQCVQKYGTEFCKKRLA from rAni s 7 was identified as the epitope recognized by mAb UA3, and is the target for over 60% of human IgE antibodies that react with O-deglycosylated nAni s 7. Conclusions: In addition to their clear value for serodiagnosis of human anisakiasis, the nature of the novel sequences and epitopes identified in the Ani s 7 allergen are of interest for a better understanding of the mechanisms operating in Anisakis-induced allergy. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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