Detection of activated basophils using flow cytometry for diagnosis in atopic patients.
BACKGROUND
human basophils release mediators of allergy after cross-linking of IgE receptors by allergens. Specific activation of basophils is detectable through flow cytometry (FCM) using an anti-CD63 fluorescein-conjugated monoclonal antibody.


OBJECTIVE
this study evaluate the detection of activated basophils by FCM in routine diagnosis of atopic diseases as regard to skin prick tests and specific immunoglobulin E antibodies.


RESULTS
whole blood from twenty patients suspected of atopy was preincubated with interleukin-3 (IL-3), then incubated with specific allergens. After staining using anti-CD63 antibodies, activated basophils were detected through FCM. IL-3-preincubation increases the spontaneous expression of CD63 even at low concentrations (0.1 ng/ml) on the basophils of 2 patients out of 20. The sensitivity and specificity of FCM were respectively 0.56 +/- 0.17 (m +/- SD) and 1.0 +/- 0.0 for the detection of dust mite-activated basophils without IL-3 preincubation, and 0.73 +/- 0.13 and 1.0 +/- 0.0 for the detention of grass pollen-activated basophils. IL-3-preincubation increased the sensitivity in a dose-dependent manner but decreased the specificity fo FCM for detecting dust mite hypersensitivity.


CONCLUSIONS
this method allow for rapid and easy detection of activated basophils from whole blood, and could be of interest for detecting allergies to non-conventional allergens such as pharmaceutical drugs.
Cozon
G
G
Unité d'Immunologie, Hôpital de la Croix-Rousse, Lyon Cedex 04, 69317, France. gcozon@rockefeller.univ-lyon1.fr
Ferrándiz
J
J
Peyramond
D
D
Brunet
J
J
D000485:Allergens / Q000276:immunology; D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D039741:Antigens, Dermatophagoides; D001491:Basophils / Q000276:immunology*; D002469:Cell Separation; D005434:Flow Cytometry / Q000379:methods*; D006023:Glycoproteins / Q000276:immunology; D006801:Humans; D006967:Hypersensitivity / Q000175:diagnosis*; D006969:Hypersensitivity, Immediate / Q000175:diagnosis; D007073:Immunoglobulin E / Q000032:analysis; D007377:Interleukin-3; D008925:Mites / Q000276:immunology; D010980:Platelet Membrane Glycoproteins / Q000032:analysis; D012680:Sensitivity and Specificity; D060149:Tetraspanin 30
D000485:Allergens; D015703:Antigens, CD; D039741:Antigens, Dermatophagoides; C554586:CD63 protein, human; D006023:Glycoproteins; D007377:Interleukin-3; D010980:Platelet Membrane Glycoproteins; D060149:Tetraspanin 30; D007073:Immunoglobulin E