Basophil CD63 expression assay on highly sensitized atopic donor leucocytes - A useful method in chronic autoimmune urticaria

E. Gyimesi, S. Sipka, K. Dankó, E. Kiss, B. Hídvégi, M. Gál, J. Hunyadi, B. Irinyi, A. Szegedi

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Abstract

Background: The autoimmune subclass of chronic idiopathic urticaria (CU) has been characterized by the occurrence of biologically relevant IgG antibodies against the IgE molecule or the α chain of the high-affinity Fcε receptor (FcεRIα) on basophils and mast cells. These antibodies are usually detected by autologous serum skin testing and confirmed by histamine release studies, immunoblotting, or enzyme-linked immunosorbent assay, but not always. Objectives: To detect autoantibodies to the FcεRIα in sera of CU patients by a modified serum-induced basophil activation test measured by flow cytometry (FCM) and to evaluate the relationship between the in vitro functional test, the autologous serum skin test (ASST), and the serum levels of IgE, eosinophil cationic protein (ECP) and antithyroid antibodies. Methods: Sera of 30 patients with CU and 26 patients with systemic autoimmune diseases (systemic lupus erythematosus, dermatomyositis) were tested for CD63 activation marker expression on basophils by FCM. Leucocytes from two highly sensitized atopic donors (DA1. DA2) and one non-atopic donor (D NA) were incubated with patients' sera and double-labelled with anti-IgE and anti-CD63 antibodies. Subsequently, the percentage of CD63-expressing basophils was determined by using FCM. In all CU patients an ASST was carried out and the serum IgE, and ECP levels and antithyroid antibodies were evaluated. Results: Twelve patients had a positive ASST and 14 patients a positive CD63 expression assay. There was a strong correlation between the ASST and CD63 assay. Sera from patients with systemic autoimmune diseases did not raise positive CD63 expression on basophils. There was a moderate negative correlation between the occurrence of atopic serum markers (IgE, ECP) and the ability of sera to induce CD63 expression on basophil cells of DA2 (P <0.05). The female sex was preponderant and antithyroid antibodies were more frequent. Conclusions: Our new technical observation demonstrates that basophils of highly sensitized atopic donors can be successfully used without priming with IL-3 for the in-vitro flow cytofluorimetric diagnosis of CU. With this investigation the characterization of the autoimmune origin of CU is based on an objective in vitro technique.

Original languageEnglish
Pages (from-to)388-396
Number of pages9
JournalBritish Journal of Dermatology
Volume151
Issue number2
DOIs
Publication statusPublished - Aug 2004

Fingerprint

Basophils
Urticaria
Leukocytes
Tissue Donors
Serum
Eosinophil Cationic Protein
Skin Tests
Immunoglobulin E
Antibodies
Flow Cytometry
Autoimmune Diseases
Dermatomyositis
Fc Receptors
Histamine Release
Interleukin-3
Immunoblotting
Mast Cells
Systemic Lupus Erythematosus
Autoantibodies
Anti-Idiotypic Antibodies

Keywords

  • Autologous serum skin test
  • Basophil activation
  • CD63 expression
  • Chronic urticaria
  • Flow cytometry

ASJC Scopus subject areas

  • Dermatology

Cite this

@article{5564465996f84f43b95b7196c52fa6e2,
title = "Basophil CD63 expression assay on highly sensitized atopic donor leucocytes - A useful method in chronic autoimmune urticaria",
abstract = "Background: The autoimmune subclass of chronic idiopathic urticaria (CU) has been characterized by the occurrence of biologically relevant IgG antibodies against the IgE molecule or the α chain of the high-affinity Fcε receptor (FcεRIα) on basophils and mast cells. These antibodies are usually detected by autologous serum skin testing and confirmed by histamine release studies, immunoblotting, or enzyme-linked immunosorbent assay, but not always. Objectives: To detect autoantibodies to the FcεRIα in sera of CU patients by a modified serum-induced basophil activation test measured by flow cytometry (FCM) and to evaluate the relationship between the in vitro functional test, the autologous serum skin test (ASST), and the serum levels of IgE, eosinophil cationic protein (ECP) and antithyroid antibodies. Methods: Sera of 30 patients with CU and 26 patients with systemic autoimmune diseases (systemic lupus erythematosus, dermatomyositis) were tested for CD63 activation marker expression on basophils by FCM. Leucocytes from two highly sensitized atopic donors (DA1. DA2) and one non-atopic donor (D NA) were incubated with patients' sera and double-labelled with anti-IgE and anti-CD63 antibodies. Subsequently, the percentage of CD63-expressing basophils was determined by using FCM. In all CU patients an ASST was carried out and the serum IgE, and ECP levels and antithyroid antibodies were evaluated. Results: Twelve patients had a positive ASST and 14 patients a positive CD63 expression assay. There was a strong correlation between the ASST and CD63 assay. Sera from patients with systemic autoimmune diseases did not raise positive CD63 expression on basophils. There was a moderate negative correlation between the occurrence of atopic serum markers (IgE, ECP) and the ability of sera to induce CD63 expression on basophil cells of DA2 (P <0.05). The female sex was preponderant and antithyroid antibodies were more frequent. Conclusions: Our new technical observation demonstrates that basophils of highly sensitized atopic donors can be successfully used without priming with IL-3 for the in-vitro flow cytofluorimetric diagnosis of CU. With this investigation the characterization of the autoimmune origin of CU is based on an objective in vitro technique.",
keywords = "Autologous serum skin test, Basophil activation, CD63 expression, Chronic urticaria, Flow cytometry",
author = "E. Gyimesi and S. Sipka and K. Dank{\'o} and E. Kiss and B. H{\'i}dv{\'e}gi and M. G{\'a}l and J. Hunyadi and B. Irinyi and A. Szegedi",
year = "2004",
month = "8",
doi = "10.1111/j.1365-2133.2004.06042.x",
language = "English",
volume = "151",
pages = "388--396",
journal = "British Journal of Dermatology",
issn = "0007-0963",
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TY - JOUR

T1 - Basophil CD63 expression assay on highly sensitized atopic donor leucocytes - A useful method in chronic autoimmune urticaria

AU - Gyimesi, E.

AU - Sipka, S.

AU - Dankó, K.

AU - Kiss, E.

AU - Hídvégi, B.

AU - Gál, M.

AU - Hunyadi, J.

AU - Irinyi, B.

AU - Szegedi, A.

PY - 2004/8

Y1 - 2004/8

N2 - Background: The autoimmune subclass of chronic idiopathic urticaria (CU) has been characterized by the occurrence of biologically relevant IgG antibodies against the IgE molecule or the α chain of the high-affinity Fcε receptor (FcεRIα) on basophils and mast cells. These antibodies are usually detected by autologous serum skin testing and confirmed by histamine release studies, immunoblotting, or enzyme-linked immunosorbent assay, but not always. Objectives: To detect autoantibodies to the FcεRIα in sera of CU patients by a modified serum-induced basophil activation test measured by flow cytometry (FCM) and to evaluate the relationship between the in vitro functional test, the autologous serum skin test (ASST), and the serum levels of IgE, eosinophil cationic protein (ECP) and antithyroid antibodies. Methods: Sera of 30 patients with CU and 26 patients with systemic autoimmune diseases (systemic lupus erythematosus, dermatomyositis) were tested for CD63 activation marker expression on basophils by FCM. Leucocytes from two highly sensitized atopic donors (DA1. DA2) and one non-atopic donor (D NA) were incubated with patients' sera and double-labelled with anti-IgE and anti-CD63 antibodies. Subsequently, the percentage of CD63-expressing basophils was determined by using FCM. In all CU patients an ASST was carried out and the serum IgE, and ECP levels and antithyroid antibodies were evaluated. Results: Twelve patients had a positive ASST and 14 patients a positive CD63 expression assay. There was a strong correlation between the ASST and CD63 assay. Sera from patients with systemic autoimmune diseases did not raise positive CD63 expression on basophils. There was a moderate negative correlation between the occurrence of atopic serum markers (IgE, ECP) and the ability of sera to induce CD63 expression on basophil cells of DA2 (P <0.05). The female sex was preponderant and antithyroid antibodies were more frequent. Conclusions: Our new technical observation demonstrates that basophils of highly sensitized atopic donors can be successfully used without priming with IL-3 for the in-vitro flow cytofluorimetric diagnosis of CU. With this investigation the characterization of the autoimmune origin of CU is based on an objective in vitro technique.

AB - Background: The autoimmune subclass of chronic idiopathic urticaria (CU) has been characterized by the occurrence of biologically relevant IgG antibodies against the IgE molecule or the α chain of the high-affinity Fcε receptor (FcεRIα) on basophils and mast cells. These antibodies are usually detected by autologous serum skin testing and confirmed by histamine release studies, immunoblotting, or enzyme-linked immunosorbent assay, but not always. Objectives: To detect autoantibodies to the FcεRIα in sera of CU patients by a modified serum-induced basophil activation test measured by flow cytometry (FCM) and to evaluate the relationship between the in vitro functional test, the autologous serum skin test (ASST), and the serum levels of IgE, eosinophil cationic protein (ECP) and antithyroid antibodies. Methods: Sera of 30 patients with CU and 26 patients with systemic autoimmune diseases (systemic lupus erythematosus, dermatomyositis) were tested for CD63 activation marker expression on basophils by FCM. Leucocytes from two highly sensitized atopic donors (DA1. DA2) and one non-atopic donor (D NA) were incubated with patients' sera and double-labelled with anti-IgE and anti-CD63 antibodies. Subsequently, the percentage of CD63-expressing basophils was determined by using FCM. In all CU patients an ASST was carried out and the serum IgE, and ECP levels and antithyroid antibodies were evaluated. Results: Twelve patients had a positive ASST and 14 patients a positive CD63 expression assay. There was a strong correlation between the ASST and CD63 assay. Sera from patients with systemic autoimmune diseases did not raise positive CD63 expression on basophils. There was a moderate negative correlation between the occurrence of atopic serum markers (IgE, ECP) and the ability of sera to induce CD63 expression on basophil cells of DA2 (P <0.05). The female sex was preponderant and antithyroid antibodies were more frequent. Conclusions: Our new technical observation demonstrates that basophils of highly sensitized atopic donors can be successfully used without priming with IL-3 for the in-vitro flow cytofluorimetric diagnosis of CU. With this investigation the characterization of the autoimmune origin of CU is based on an objective in vitro technique.

KW - Autologous serum skin test

KW - Basophil activation

KW - CD63 expression

KW - Chronic urticaria

KW - Flow cytometry

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U2 - 10.1111/j.1365-2133.2004.06042.x

DO - 10.1111/j.1365-2133.2004.06042.x

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EP - 396

JO - British Journal of Dermatology

JF - British Journal of Dermatology

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