Flow cytometry versus histamine release analysis of in vitro basophil degranulation in allergy to Hymenoptera venom.
BACKGROUND
Flow cytometry (FCM) has been proposed for specific allergy in vitro testing. We investigated its biological significance for allergy to Hymenoptera venoms and compared it with the routinely performed basophil histamine release test (HRT).


METHODS
Blood samples from 26 allergic and 8 nonallergic donors were incubated with venom at serial concentrations. Basophils were analyzed with anti-CD45-PE-Cyanin 5, Anti-IgE-FITC, and Anti-CD63-Phycoerythrine. HRT was measured by radioimmunoassay.


RESULTS
FCM was as convenient as HRT for measuring basophil reactivity in at least 87% of allergic and 75% of nonallergic subjects. CD63 outer expression was specifically induced in 91% of releaser subjects (86% on HRT) and in 1 of 10 tests in nonallergic donors, or one of six tests (16% on HRT) in allergic patients tested with an irrelevant allergen. Both methods were concordant in 85.7% of the tests. The three discordant patients had low-grade reactions and borderline biological responses on FCM (n = 2) or HRT (n = 1).


CONCLUSIONS
The dynamic, physiologic significance of CD63, the dose-response curve, and dependency on ethylene-diaminetetra acetic acid suggested that both tests reflect the same mechanism.
Lambert
C
C
Immunology Laboratory, University of St Etienne, St Etienne, France. lambert@chu-st-stienne.fr
Guilloux
L
L
Dzviga
C
C
Gourgaud-Massias
C
C
Genin
C
C
D000293:Adolescent; D000328:Adult; D000368:Aged; D000485:Allergens / Q000276:immunology; D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D015549:Basophil Degranulation Test; D001491:Basophils / Q000276:immunology / Q000378:metabolism; D001514:Bee Venoms / Q000276:immunology*; D002648:Child; D000939:Epitopes; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006636:Histamine Release / Q000276:immunology*; D006801:Humans; D006927:Hymenoptera; D006967:Hypersensitivity / Q000175:diagnosis*; D007073:Immunoglobulin E / Q000032:analysis; D066298:In Vitro Techniques; D017493:Leukocyte Common Antigens / Q000032:analysis; D008297:Male; D008875:Middle Aged; D010980:Platelet Membrane Glycoproteins / Q000032:analysis; D060149:Tetraspanin 30; D014862:Wasp Venoms / Q000276:immunology*
D000485:Allergens; D015703:Antigens, CD; D001514:Bee Venoms; C554586:CD63 protein, human; D000939:Epitopes; D010980:Platelet Membrane Glycoproteins; D060149:Tetraspanin 30; D014862:Wasp Venoms; D007073:Immunoglobulin E; D017493:Leukocyte Common Antigens