Recombinant allergens promote expression of CD203c on basophils in sensitized individuals

doi:10.1067/mai.2002.125257

Journal of Allergy and Clinical Immunology

Volume 110, Issue 1, July 2002, Pages 102-109

https://doi.org/10.1067/mai.2002.125257 Get rights and content

Abstract

Background: Traditionally, the diagnosis of type I allergies is based on clinical data, skin test results, and laboratory test results with allergen extracts. During the past few years, several attempts have been made to refine diagnostic assays in clinical allergy by introducing recombinant allergens and novel markers of IgE-dependent cell activation. Objectives: We have identified the ectoenzyme CD203c as a novel basophil antigen that is upregulated on IgE receptor cross-linkage. In this study we applied CD203c and a panel of recombinant allergens to establish a novel basophil test that allows for a reliable quantification of IgE-dependent responses at the effector cell level. Methods: Patients allergic to birch (Bet v 1, n = 15; Bet v 2, n = 8) and grass (Phl p 1, n = 15; Phl p 2, n = 10; Phl p 5, n = 14) pollen allergens, as well as 10 nonallergic donors, were examined. Basophils were exposed to various concentrations of recombinant allergens for 15 minutes and then examined for expression of CD203c by means of flow cytometry. CD203c upregulation was correlated with the increase in CD63. Results: Exposure to recombinant allergens resulted in a dose-dependent increase in expression of CD203c on peripheral blood basophils in sensitized individuals, whereas no increase was seen in healthy control subjects. The effects of the recombinant allergens on CD203c expression were also time dependent. There was a good correlation between allergen-induced upregulation of CD203c and upregulation of CD63 (R = 0.76). Conclusion: Flow cytometric quantitation of CD203c on blood basophils exposed to recombinant allergens is a useful approach to determine the allergic state in sensitized individuals and represents a basis for a sensitive novel allergy test. (J Allergy Clin Immunol 2002;110:102-9.)

Section snippets

Patients

Twenty patients (7 female and 13 male patients) allergic to one or more grass pollen or birch pollen allergens were examined. Patients were allergic to Bet v 1 (n = 15), Bet v 2 (n = 8), Phl p 1 (n = 15), Phl p 2 (n = 10), or Phl p 5 (n = 14). Allergy diagnosis was based on skin tests, specific IgE measurements (RAST), and clinical history. Levels of IgE were expressed as kilounits of antigen per liter. The patients' characteristics are shown in Table I. The control group consisted of 10

Recombinant allergens promote expression of CD203c on basophils in sensitized individuals in a dose-dependent manner

Recent data suggest that CD203c is a specific marker for human basophils and is upregulated in response to FcϵRI cross-linkage.¹⁹ In this study, unstimulated basophils obtained from patients with grass or birch pollen allergy expressed detectable levels of CD203c similar to those in normal basophils, whereas other blood leukocytes appeared to be CD203c negative. The specificity of CD203c for basophils in allergic donors was confirmed by sorting experiments (Fig 1).

. Morphology of CD203c⁺ and

Discussion

A number of previous studies have shown that flow cytometric quantitation of basophil activation can be used to measure IgE-dependent responses to allergens in sensitized individuals.15, 16, 17, 18, 21 Thus far, most tests have used antibodies against CD63 and crude allergen extracts.15, 16, 17, 18 In this study, we have used recombinant allergens and the novel basophil marker CD203c19, 20 to measure and quantify allergen-induced activation of basophils. Our data show that CD203c represents a

Acknowledgements

We thank Dr André van Agthoven (Immunotech) for providing phycoerythrin-labeled 97A6 mAb and Hans Semper for excellent technical assistance.

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Mast cells (MC) and basophils are effector cells of allergic reactions and display a number of activation-linked cell surface antigens. Of these antigens, however, only a few are functionally relevant and specifically expressed in these cells.
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^☆: Supported by Fonds zur Förderung der Wissenschaftlichen Forschung in Österreich (FWF) grants no. 01801, 01809, and Y078GEN.

^☆☆: Reprint requests: Peter Valent, MD, Department of Internal Medicine I, Division of Hematology and Hemostaseology, University of Vienna, Währinger Gürtel 18-20, A-1090 Vienna, Austria.

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Mechanisms of AllergyRecombinant allergens promote expression of CD203c on basophils in sensitized individuals☆,☆☆

Abstract

Section snippets

Patients

Recombinant allergens promote expression of CD203c on basophils in sensitized individuals in a dose-dependent manner

Discussion

Acknowledgements

Curr Opin Immunol

J Allergy Clin Immunol

J Biol Chem

J Invest Dermatol

J Allergy Clin Immunol

J Allergy Clin Immunol

J Allergy Clin Immunol

Blood

Blood

Biochem Biophys Res Commun

J Invest Dermatol

J Allergy Clin Immunol

Allergy and allergic diseases

Human anaphylaxis

Recombinant allergens

Allergy

Recombinant allergen molecules: tools to study effector cell activation

Immunol Rev

Mechanisms of Allergy
Recombinant allergens promote expression of CD203c on basophils in sensitized individuals☆,☆☆