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.
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.
Authors: Peter Valent; Cem Akin; Patrizia Bonadonna; Karin Hartmann; Knut Brockow; Marek Niedoszytko; Boguslaw Nedoszytko; Frank Siebenhaar; Wolfgang R Sperr; Joanna N G Oude Elberink; Joseph H Butterfield; Ivan Alvarez-Twose; Karl Sotlar; Andreas Reiter; Hanneke C Kluin-Nelemans; Olivier Hermine; Jason Gotlib; Sigurd Broesby-Olsen; Alberto Orfao; Hans-Peter Horny; Massimo Triggiani; Michel Arock; Lawrence B Schwartz; Dean D Metcalfe Journal: J Allergy Clin Immunol Pract Date: 2019-02-05
Authors: Dubravka Smiljkovic; Renata Kiss; Christian Lupinek; Gregor Hoermann; Georg Greiner; Nadine Witzeneder; Gerhard Krajnik; Franz Trautinger; Susanne Vrtala; Irene Mittermann; Michael Kundi; Bernd Jilma; Rudolf Valenta; Wolfgang R Sperr; Peter Valent Journal: J Allergy Clin Immunol Pract Date: 2020-04-26
Authors: Katharina Marth; Isabella Breyer; Margarete Focke-Tejkl; Katharina Blatt; Mohamed H Shamji; Janice Layhadi; Anna Gieras; Ines Swoboda; Domen Zafred; Walter Keller; Peter Valent; Stephen R Durham; Rudolf Valenta Journal: J Immunol Date: 2013-02-25 Impact factor: 5.422