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Histamine release-neutralization assay for sera of patients with atopic dermatitis and/or cholinergic urticaria is useful to screen type I hypersensitivity against sweat antigens

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Abstract

We previously reported that about 80 % of patients with atopic dermatitis and 60 % with cholinergic urticaria revealed type I allergy against sweat, by means of skin test against autologous sweat and/or histamine-release test for peripheral blood basophils with semi-purified sweat antigen. In this study, we developed an assay for sera to neutralize histamine-releasing activity of semi-purified sweat antigen. The semi-purified sweat antigen was pre-incubated with serially diluted sera for 30 min at 37 °C and was subjected to histamine-release activity. Histamine release-neutralization (HRN) activities were calculated by measuring the amount of histamine release from basophils in the presence or absence of semi-purified sweat antigen. Of 62 subjects, 39 showed positive histamine release (≥5 %) from their basophils in response to semi-purified sweat antigen, and sera of 34 out of 39 subjects (87.2 %) were also positive in HRN activity (≥10 %). The specificity of the HRN assay was 0.522. Moreover, HRN activities in sera were largely correlated with degrees of histamine release from peripheral blood basophils of the same donors in response to sweat antigen. To identify the substance that neutralizes histamine-release activity, we removed IgE and IgG from the sera of HRN (+) subjects by column chromatography. The HRN activities in 30 out of 42 sera were largely reduced by the removal of IgG. On the other hand, sera of four subjects lost HRN activity by the removal of IgE, suggesting that the majority of HRN (+) subjects have serum IgG against the sweat antigen as well as IgE bound to peripheral basophils. Thus, the HRN assay maybe useful for the screening of type I allergy against sweat antigen.

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Abbreviations

HRA:

Histamine-release assay

HRN:

Histamine release-neutralization

IgE:

Immunoglobulin E

IgG:

Immunoglobulin G

CAP-FEIA:

Capsulated-fluorenzymeimmunoassay

ELISA:

Enzyme-linked immunosorbent assay

ANOVA:

Analysis of variance

FACS:

Fluorescence activated cell sorter

ROC:

Receiver-operator characteristic

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Acknowledgments

We thank Ms Kazue Uchida for the expert technical assistances, and Dr Faiz Kermani for his manuscript review. This work was supported in part by Health Science Research Grants from Ministry of Health, Welfare and Labor, Japan.

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Authors and Affiliations

  1. Programs for Biomedical Research, Division of Molecular Medical Science, Department of Dermatology, Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan

    Hajime Shindo, Kaori Ishii, Yuhki Yanase, Hidenori Suzuki & Michihiro Hide

  2. Faculty of Human Science, Hiroshima Bunkyo Women’s University, 1-2-1 Kabehigasi, Asakita-ku, Hiroshima, 731-0295, Japan

    Hidenori Suzuki

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  1. Hajime Shindo
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  2. Kaori Ishii
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  3. Yuhki Yanase
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  4. Hidenori Suzuki
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  5. Michihiro Hide
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Correspondence to Michihiro Hide.

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Shindo, H., Ishii, K., Yanase, Y. et al. Histamine release-neutralization assay for sera of patients with atopic dermatitis and/or cholinergic urticaria is useful to screen type I hypersensitivity against sweat antigens. Arch Dermatol Res 304, 647–654 (2012). https://doi.org/10.1007/s00403-012-1236-2

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