Heterogeneity of IgE epitopes of vinyl sulphone reactive dye: human serum albumin that react with IgE
Article first published online: 7 JUL 2008
Clinical & Experimental Allergy
Volume 31, Issue 11, pages 1779–1786, November 2001
How to Cite
Park, J. W., Kang, D. B., Choi, S. Y., Kim, C. W., Kim, K. S. and Hong, C.-S. (2001), Heterogeneity of IgE epitopes of vinyl sulphone reactive dye: human serum albumin that react with IgE. Clinical & Experimental Allergy, 31: 1779–1786. doi: 10.1046/j.1365-2222.2001.01194.x
- Issue published online: 7 JUL 2008
- Article first published online: 7 JUL 2008
- Submitted 30 January 2001; revised 21 March 2001; accepted 30 April 2001.
- vinyl sulphone reactive dye;
- IgE epitopes;
- occupational asthma
Background Vinyl sulphone reactive dye (vRD), which consists of vinyl sulphone reactive groups and a chromogen, can elicit IgE-mediated occupational asthma (OA) by haptenation. Human serum albumin (HSA) is known as the most reliable carrier protein for the vRD, the IgE epitopes of vRD-HSA are not well characterized. In this study we evaluated the epitope of vRD-HAS-specific IgE.
Methods Two vRD (Remazole Black-GR and Remazole Orange-3R), Procion Red-MX-5B, which has a dichlorotriazine reactive group, and vinyl sulphone (VS), were haptenated to HSA, respectively. vRD-HSA was denatured by heat or mercaptoethanol treatment and the allergenicities of denatured and non-denatured vRD-HSA were compared by ELISA and IgE immunoblotting using the sera of six vRD-OA patients. vRD-HSA-specific, Procion Red-MX-5B (pRD)-HSA-specific and VS-HAS-specific IgE were also measured with ELISA and the cross-reactivity between them was evaluated with ELISA inhibition.
Results Denaturation of vRD-HSA by heat affected its allergenicity markedly in five of six sera of RD-OA. When vRD was conjugated to the pre-heated HSA, its allergenicity also disappeared or was markedly attenuated compared with the vRD-HSA in five of six sera. Mercaptoethanol treatment markedly affected the allergenicity of the RD-HSA in all six RD-OA sera. Immunoblotting from non-denatured PAGE showed strong IgE affinity to vRD-HSA but immunoblotting from denatured SDS PAGE did not show IgE affinity. Among six RD-OA patients, five and four patients had pRD-HSA-specific and VS-HSA-specific IgE, respectively. However, the vRD-HSA-specific IgE was neither inhibited by pRD-HSA nor VS-HSA
Conclusion We considered that the conformational structure of HSA would be critical for the IgE epitopes during the haptenation process and both of the chromogen and reactive groups of the vRD would contribute to the formation of IgE epitope. Our results also confirmed the heterogeneity of IgE epitopes in the RD-HSA complex.