Semi-purification of the immunoglobulin E-sweat antigen acting on mast cells and basophils in atopic dermatitis

Exp Dermatol. 2006 Apr;15(4):283-90. doi: 10.1111/j.0906-6705.2006.00404.x.

Abstract

Background: Sweating aggravates the symptoms of atopic dermatitis (AD). We have recently reported positive skin reactions and histamine release from basophils in response to autologous sweat in patients with AD.

Objective: To characterize the biochemical and immunological properties of the substance in sweat that evokes histamine release and to study the usability of the basophil-histamine release test with the sweat antigen for AD.

Methods: Sweat collected from healthy volunteers was purified using chromatographies. Serum immunoglobulin (Ig)E of four patients with AD were purified using an affinity-chromatography column with anti-IgE antibodies. The amount of semi-purified sweat antigen (138 ng protein/ml) that induced a half-maximum reaction of basophils of a patient with AD was utilized for the basophil histamine release test. The involvement of specific IgE and high-affinity IgE receptor (FcepsilonRI) in the reactions was examined using basophils of healthy volunteers, a human mast cell line (LAD2), and a rat basophilic leukemia cell line transfected with human alpha-subunit of FcepsilonRI (RBL-48).

Results: The semi-purified sweat antigen induced histamine release from the basophils of 47 of 61 (74.6%) patients with AD and four of 46 (8.7%) healthy controls. Both basophils and mast cells sensitized with the patient-derived IgE showed degranulation upon stimulation with the sweat antigen. However, no reaction was observed when cells were sensitized with myeloma IgE or the antigen was treated with proteases.

Conclusion: The semi-purified standardized sweat antigen consists of a protein that induces degranulation of basophils and mast cells via antigen-specific IgE and FcepsilonRI in patients with AD.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Animals
  • Basophils / immunology*
  • Cell Line
  • Cell Line, Tumor
  • Child
  • Child, Preschool
  • Chromatography, Affinity
  • Dermatitis, Atopic / immunology*
  • Dermatitis, Atopic / physiopathology
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Histamine Release
  • Humans
  • Immunoglobulin E / immunology
  • Immunoglobulin E / isolation & purification*
  • Infant
  • Male
  • Mast Cells / immunology*
  • Middle Aged
  • Receptors, IgE / physiology
  • Sensitivity and Specificity
  • Sweat / immunology*
  • beta-N-Acetylhexosaminidases / metabolism

Substances

  • Receptors, IgE
  • Immunoglobulin E
  • beta-N-Acetylhexosaminidases