Diagnosis of venom allergy by flow cytometry. Correlation with clinical history, skin tests, specific IgE, histamine and leukotriene C4 release.
BACKGROUND
Potent allergens such as hymenoptera venoms are capable of inducing severe and life threatening clinical reactions. Percentage of false negative results obtained by the usual diagnostical methods is comprised between 10 and 25%.


OBJECTIVE
Evaluation of the sensitivity and the specificity of cellular tests and particularly evaluation of a new flow cytometric method.


METHODS
Forty-five allergic patients having experienced a local, a systemic reaction or an anaphylactic shock and 10 controls having undergone hymenoptera stings without clinical reactions were selected on the basis of the clinical history, skin tests and specific IgE. Three cellular tests were performed on the same cell suspensions and in the presence of 2 ng/mL of rIL3: histamine release (RIA), leukotriene C4 release (ELISA) and basophil activation test (flow cytometry after double anti-IgE FITC, anti-CD63 PE labelling).


RESULTS
As compared to the clinical history, sensitivities of skin tests, specific IgE, flow cytometry, histamine release and leukotriene release were, respectively; 85%, 88%, 100%, 89% and 100%. Flow cytometric analysis of basophil activation showed a significant decrease of the mean fluorescence density and number of IgE positive cells and a significant increase of the number of CD63 positive cells. The 10 controls tested by flow cytometry were negative.


CONCLUSIONS
As compared to the clinical history and to the other parameters tested here, flow cytometry showed a high sensitivity and a high specificity. The excellent correlation observed between this method and the other cellular tests such as histamine and leukotriene release are in favour of the specificity of flow cytomery and in favour of the use of this method for venom allergy diagnosis.
Unité d'immuno-allergologie, laboratoire Pasteur-Cerba, Cergy-Pontoise, France.
J
J
Sainte-Laudy
A
A
Sabbah
M
M
Drouet
M G
MG
Lauret
M
M
Loiry
D000293:Adolescent; D000328:Adult; D000486:Allergy and Immunology; D000818:Animals; D015703:Antigens, CD / Q000276:immunology / Q000378:metabolism; D001180:Arthropod Venoms / Q000009:adverse effects* / Q000276:immunology; D001491:Basophils / Q000276:immunology / Q000378:metabolism*; D015415:Biomarkers / Q000032:analysis; D002648:Child; D005434:Flow Cytometry; D006636:Histamine Release; D006801:Humans; D006927:Hymenoptera; D006967:Hypersensitivity / Q000175:diagnosis* / Q000276:immunology; D007073:Immunoglobulin E / Q000276:immunology / Q000378:metabolism*; D007299:Insect Bites and Stings / Q000276:immunology; D007962:Leukocytes / Q000276:immunology / Q000378:metabolism; D017997:Leukotriene C4 / Q000276:immunology / Q000378:metabolism*; D008875:Middle Aged; D010980:Platelet Membrane Glycoproteins / Q000276:immunology / Q000378:metabolism; D012680:Sensitivity and Specificity; D012882:Skin Tests; D013223:Statistics as Topic; D060149:Tetraspanin 30
D015703:Antigens, CD; D001180:Arthropod Venoms; D015415:Biomarkers; C554586:CD63 protein, human; D010980:Platelet Membrane Glycoproteins; D060149:Tetraspanin 30; D017997:Leukotriene C4; D007073:Immunoglobulin E