Naslov
Food allergy diagnosis: in vitro method for measurement of basophil activation and/or degranulation in children with food allergies

Autori
Živković, Jelena ; Polančec, Denis ; Zenić, Lucija ; Bulat Lokas, Sandra ; Turkalj, Mirjana

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
EAACI Congress 2017 : abstracts / - , 2017, Xx-xx

Skup
EAACI Congress 2017

Mjesto i datum
Helsinki, Finska, 17.06.2017. - 21.06.2017

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
BAT, food allergy, IgE

Sažetak
Basophil activation test (BAT) is a functional test that reflects the ex vivo IgE- mediated cell response induced by certain allergens and seems to be more specific and more sensitive for food allergy testing than other available in vitro tests. It represents a new tool developed to monitor basophil activation upon allergen challenge by detecting the expression of membrane surface markers (CD63, CD203c) by flow cytometry. Although some studies claimed there are advantages of CD63 over CD203c, several studies addressed the role of CD203c as the most promising new activation marker for flow-cytometry based allergy diagnosis. To introduce BAT to the daily routine and help clinicians in cases where other diagnostic procedures give ambiguous results, i.e. show discrepancies between skin prick test, specific IgE and patient clinical history. To test CD203c as a basophil activation marker and to determine its correlation with CD63. Peripheral blood was obtained from children allergic to food. Samples were prepared using the FlowCAST® (Bühlmann, Switzerland) protocol with addition of antibodies for CD45 and CD203c. Acquistion of the samples was performed using the Navios flow cytometer (Beckman Coulter, USA), while data were analysed using the FlowLogic™ software (Inivai Technologies, Australia). For both markers fraction of activated basophils and the "mean fluorescence intensity" (MFI) as a measure of expression was determined in parallel. The simultaneous use of four different antibodies allows a better segregation and a precise measurement of basophil activation and/or degranulation. Also, the MFI for CD203c significantly differed among samples treated with different allergen concentrations although the percentage of activated basophils estimated by CD63 expression did not differ between those samples, i.e. the differences in CD203c expression are measurable during non-degranulating stimulation of basophils. BAT is a reliable and useful functional test in cases demonstrating inconclusive diagnostic results. The use of anti- CD203c antibody, as an additional identification and activation marker, has been shown useful for a better resolution of the activated/non- degranulated and activated/degranulated basophils. The use of CD203c improved both the sensitivity and the specificity of BAT, resulting in an enhancement of its accuracy in the diagnosis of food allergies.

Izvorni jezik
Engleski

Znanstvena područja
Kliničke medicinske znanosti

POVEZANOST RADA