Der p 2 isoallergens have different allergenicity, and quantification with 2-site ELISA using monoclonal antibodies is influenced by the isoallergens

J. W. Park, K. S. Kim, H. S. Jin, C. W. Kim, D. B. Kang, S. Y. Choi, T. S. Yong, S. H. Oh, C. S. Hong

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Abstract

Background: Der p 2 isoallergens have been reported and the possibility of different allergenicity has also been suggested. In addition, the quantification with 2-site ELISA may be affected by the isoallergens. Objectives: Two different recombinant Der p 2 (rDer p 2) isoallergens were compared in terms of human IgE responses and the reliability of quantification of them with two-site ELISA kits which use monoclonal antibodies (mAbs) as capture and detection of Der p 2. Methods: Seven different Der p 2 cDNA from the cultured Dermatophagoides pteronyssinus (DP) were cloned and polymorphism in nine amino acid residues was found. Two different recombinant isoallergens (rDer p 2A and rDer p 2B) were expressed and compared to their human IgE immune responses by ELISA and the ELISA inhibition test with 23 sera of DP-allergic patients. The reliability of quantification of two different available 2-site ELISA kits, which used mAbs for capture and detection of Der p 2, was evaluated. Results: The ELISA optical density of rDer p 2B-specific IgE (slgE) was higher than that of rDer p 2A (P < 0.001). The ELISA inhibition curve of rDer p 2B slgE in pool I sera (n = 5; high slgE both to rDer p 2A and rDer p 2B) did not show any differences in the 50% inhibition concentration and maximum inhibitory percentage of rDer p 2A and rDer p 2B slgE. However, with pool II sera (n = 5; markedly higher slgE to rDer p 2B than rDer p 2A), the 50% inhibitory concentrations (10 μg/mL vs. 40 ng/mL) and maximum inhibitory percentage (61% vs. 99%) of rDer p 2B slgE with the two recombinant isoallergens were quite different, rDer p 2B could be quantified with two different 2-site ELISA kits, but rDer p 2A was detected by only one kit. Conclusion: We conclude that isoallergens of Der p 2 may have different IgE immune responses. Quantification of Der p 2 with 2-site ELISA kits that adopted mAbs, might be affected by the prevalent form of the isoallergens in reservoir dust.

Original languageEnglish
Pages (from-to)1042-1047
Number of pages6
JournalClinical and Experimental Allergy
Volume32
Issue number7
DOIs
Publication statusPublished - 2002 Jul 27

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All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

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