Adducts between nucleophilic amino acids and hexahydrophthalic anhydride, a structure inducing both types I and IV allergy

Abstract

Haptens causing type I allergy have been shown to predominantly form lysine adducts in the carrier protein, while many haptens giving rise to type IV allergy preferentially form adducts with cysteine residues. Hexahydrophthalic anhydride derivatives are strong sensitizers capable of inducing allergic rhinitis, asthma and urticaria (type I allergy) and allergic contact dermatitis (type IV allergy). The ability of hexahydrophthalic anhydride (HHPA) to form adducts with nucleophilic amino acids and a model peptide in vitro is presented. Adduct formation was monitored by high-performance liquid chromatography with ultraviolet light/vis detection (LC-UV/vis) and high-performance liquid chromatography with mass spectrometric detection (LC/MS). The characterization was obtained by nuclear magnetic resonance spectroscopy (NMR) and mass spectrometry (MS and MS/MS). It was found that HHPA formed adducts with N^α-acetylated lysine and cysteine and the non-acetylated α-amino group of proline and, to some extent, also with other nucleophilic amino acids. The adducts with lysine and proline were chemically stable. Addition of one HHPA to a model carrier peptide with all important nucleophilic amino acid residues showed N-terminal proline to be the major site of reaction. The addition of a second hapten gave a lysine adduct, but a minor cysteine adduct was also found. The cysteine–HHPA adducts were shown to be chemically unstable and participated in further reactions with lysine forming lysine–HHPA adducts. The results will be useful for understanding the formation of HHPA–protein adducts with the capability of being markers of exposure, and also to a deeper understanding of the chemical structures causing types I and IV allergy.

Keywords: :

• Adduct
• allergy
• amino acids
• antigen
• hapten
• hexahydrophthalic anhydride

Acknowledgements

The investigation was supported by grants from the Swedish Research Council, the Swedish Council for Working Life and Social Research, the Swedish Foundation for Health Care and Allergy Research, the Alfred Österlund Foundation for Scientific Research, the Edvard Welander Foundation, and the Medical Faculty at Lund University.