Hymenoptera-venom-induced upregulation of the basophil activation marker ecto-nucleotide pyrophosphatase/phosphodiesterase 3 in sensitized individuals.
BACKGROUND
Bee and wasp venoms are potent allergens capable of inducing severe clinical reactions. To detect immediate-type hypersensitivity to these allergens, a rapid in vitro test was developed that relies on the upregulation of ecto-nucleotide pyrophosphatase/phosphodiesterase 3 (E-NPP3) on activated basophils.


METHODS
Blood basophils of 13 healthy donors and 22 patients allergic to bee or wasp venom were analyzed for E-NPP3 (CD203c) expression using monoclonal antibody 97A6. Basophils were analyzed by flow cytometry after activation with anti-IgE antibody or allergen. Venom-induced E-NPP3 upregulation on basophils was compared with diagnostic parameters, including skin tests and assessment of specific IgE. In selected samples, the increase in E-NPP3 expression on activated basophils was compared with histamine release and CD63 upregulation.


RESULTS
In 20/22 patients sensitized to wasp or bee venom, E-NPP3 expression on basophils was upregulated in response to activation by allergen or anti-IgE. The maximum increase in E-NPP3 expression (above ten times of baseline) was achieved after 15 min of stimulation with 1 microg/ml of allergen or anti-IgE antibody. Sensitized individuals who failed to upregulate E-NPP3 in response to IgE receptor cross-linking also failed to induce histamine release and CD63 upregulation.


CONCLUSIONS
Flow cytometric determination of hymenoptera-venom-induced upregulation of E-NPP3 is a novel in vitro test to identify sensitized individuals.
Platz
I J
IJ
University of Tübingen, Department of Internal Medicine II, Division of Hematology and Immunology, Tübingen, Germany.
Binder
M
M
Marxer
A
A
Lischka
G
G
Valent
P
P
Bühring
H J
HJ
D000328:Adult; D000368:Aged; D000485:Allergens / Q000009:adverse effects / Q000276:immunology*; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001491:Basophils / Q000276:immunology* / Q000378:metabolism; D001514:Bee Venoms / Q000009:adverse effects / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D006636:Histamine Release; D006801:Humans; D006927:Hymenoptera / Q000276:immunology; D006969:Hypersensitivity, Immediate / Q000276:immunology; D007073:Immunoglobulin E / Q000032:analysis; D008297:Male; D008875:Middle Aged; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism; D011755:Pyrophosphatases / Q000378:metabolism*; D060149:Tetraspanin 30; D015854:Up-Regulation; D014862:Wasp Venoms / Q000009:adverse effects / Q000276:immunology*
D000485:Allergens; D015703:Antigens, CD; D001514:Bee Venoms; C554586:CD63 protein, human; C427944:ENPP3 protein, human; D010980:Platelet Membrane Glycoproteins; D060149:Tetraspanin 30; D014862:Wasp Venoms; D007073:Immunoglobulin E; D010727:Phosphoric Diester Hydrolases; D011755:Pyrophosphatases