Individual hymenoptera venom compounds induce upregulation of the basophil activation marker ectonucleotide pyrophosphatase/phosphodiesterase 3 (CD203c) in sensitized patients.
BACKGROUND
Bee and wasp venom extracts contain potent allergens capable of inducing severe clinical reactions. To analyze immediate-type hypersensitivity to defined hymenoptera venom components, a recently developed in vitro test was applied that is based on the upregulation of CD203c expression on basophils.


METHODS
CD203c expression on blood basophils of 9 healthy donors and 39 patients allergic to bee and/or wasp venom was analyzed by flow cytometry before and after activation with the purified bee venom allergens phospholipase A2 (Api m 1), hyaluronidase (Api m 2) and melittin (Api m 4), or the purified wasp venom allergens phospholipase A1 (Ves v 1), hyaluronidase (Ves v 2) and the recombinant antigen 5 (Ves v 5). Venom-induced CD203c upregulation on basophils was compared with skin tests and assessment of specific IgE. Basophils of nonresponders were preincubated with 10 ng/ml interleukin-3 (IL-3) prior to allergen stimulation.


RESULTS
CD203c upregulation on basophils was induced by defined hymenoptera venom components in 35/39 patients with a diagnosed allergy to wasp and/or bee venom. Twenty-seven of the 34 tested patients with wasp allergy showed CD203c upregulation in response to Ves v 5, 26 of these patients also reacted with Ves v 2 and 17 with Ves v 1. Nine of 13 patients with bee allergy reacted with Api m 1, 13 individuals with Api m 2 and none of these patients with the minor allergen Api m 4. A diagnosed wasp allergy could also be confirmed in the prestimulated basophils (IL-3) of 2 nonresponder individuals who failed to upregulate CD203c in response to IgE receptor cross-linking prior to culture with IL-3.


CONCLUSIONS
Flow-cytometric determination of CD203c upregulation on basophils activated by molecularly defined allergens is a powerful method to identify the precise allergen reactivity in sensitized individuals.
Binder
Marc
M
Department of Internal Medicine II, Division of Hematology and Immunology, University of Tübingen, Germany.
Fierlbeck
Gerhard
G
King
Te iaoP
Ti
Valent
Peter
P
Bühring
Hans-Jörg
HJ
D000293:Adolescent; D000328:Adult; D000368:Aged; D000485:Allergens / Q000276:immunology; D000818:Animals; D001491:Basophils / Q000276:immunology*; D001514:Bee Venoms / Q000276:immunology*; D001516:Bees / Q000276:immunology; D002648:Child; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D006967:Hypersensitivity / Q000175:diagnosis / Q000276:immunology*; D007073:Immunoglobulin E / Q000276:immunology; D007377:Interleukin-3 / Q000276:immunology; D008297:Male; D008875:Middle Aged; D010727:Phosphoric Diester Hydrolases / Q000096:biosynthesis / Q000276:immunology*; D011755:Pyrophosphatases / Q000096:biosynthesis / Q000276:immunology*; D015854:Up-Regulation / Q000276:immunology; D014862:Wasp Venoms / Q000276:immunology*; D014863:Wasps / Q000276:immunology
D000485:Allergens; D001514:Bee Venoms; C427944:ENPP3 protein, human; D007377:Interleukin-3; D014862:Wasp Venoms; D007073:Immunoglobulin E; D010727:Phosphoric Diester Hydrolases; D011755:Pyrophosphatases