scholarly journals The basophil activation test differentiates between patients with wheat‐dependent exercise‐induced anaphylaxis and control subjects using gluten and isolated gluten protein types

2021 ◽  
Vol 11 (6) ◽  
Author(s):  
Angelika Miriam Gabler ◽  
Julia Gebhard ◽  
Bernadette Eberlein ◽  
Tilo Biedermann ◽  
Katharina Anne Scherf ◽  
...  
Keyword(s):  
Basophil Activation Test ◽  
Basophil Activation ◽  
Gluten Protein ◽  
Activation Test ◽  
Control Subjects ◽  
Exercise Induced ◽  
And Control

Developing an in Vitro Functional Model of Allergic Transfusion Reactions

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 3426-3426
Author(s):  
William Savage ◽  
Aaron A. R. Tobian ◽  
Jessica Savage ◽  
Chris Thoburn ◽  
Paul M. Ness ◽  
...  
Keyword(s):  
Basophil Activation Test ◽  
Positive Control ◽  
Basophil Activation ◽  
Clinical Severity ◽  
Blue Staining ◽  
Activation Test ◽  
And Control ◽  
Transfusion Reactions ◽  
Positive Controls

Abstract Abstract 3426 Background: Allergic transfusion reactions (ATRs) occur in ∼2–3% of platelet transfusions, but their etiology remains largely unclear. Both donor and recipient factors have been shown to contribute to ATR risk. An in vitro model of ATRs would enable mechanistic studies. Objective: To develop a basophil activation test as a model of ATRs to apheresis platelets (AP) by measuring histamine release (HR) and leukotriene C4 (LTC4) production after exposing basophils to AP supernatants. Methods: Basophil-enriched suspensions (∼5–15% basophils) from healthy donors were prepared using double-Percoll density centrifugation. After washing and counting basophil numbers using Alcian blue staining, cells were resuspended in buffer containing 2 ng/mL IL-3 and 5mM Ca++/Mg++ such that 20–30×104 basophils were added in 0.025mL aliquots. IL-3 is a potent basophil priming agent and was used to model the increased recipient susceptibility to allergic reactions that has been observed clinically. Reaction tubes (1.5 mL) were set-up containing 0.025mL of 2× (twice the final conc.) of ATR/control supernatant (final titer 1:2–1:8), negative (buffer alone), and positive control reagents (100 ng/mL anti-IgE antibody, 50 ng/mL C5a, 1 μM FMLP). Cells and reaction tubes were pre-warmed separately to 37°C for 15' before aliquoting 0.025mL cell suspension to reaction tubes for a final volume of 0.050mL. After 30′ incubation at 37°C, 0.950mL of cold buffer (without IL-3 or Ca++/Mg++) was added to each reaction tube. Tubes were centrifuged 10" at ∼300g (4°C) before removing 0.900mL for histamine detection by automated fluorimetry and LTC4 detection by ELISA. AP supernatants that were (n=10) or were not (n=10) associated with ATRs were tested. HR values were determined as a percentage of the total histamine content by lysis with perchloric acid (1.6%). Results used in analysis are net HR or LTC4 production (AP sample minus IL-3 primed negative control) normalized to HR or LTC4 production observed with positive control. Results: IL-3 priming of basophils was necessary in order to evoke a response after exposure to AP supernatant. HR in response to AP incubation with unprimed basophils was <3% of positive controls. Maximal HR and LTC4 production in response to AP supernatants occurred at a final titer of 1:4 using IL-3 primed basophils. HR and LTC4 production within individual reactions were correlated (Spearman rho=0.83, P<0.0001). Mean net HR among all AP supernatant samples was 55±28%, 60±49%, and 53±29% of c5a, anti-IgE, and FMLP positive controls, respectively. LTC4 production was 19±26%, 14±22%, and 15±20% of c5a, anti-IgE, and FMLP positive controls. No statistically significant differences were observed in HR or LTC4 production when ATR and control groups were compared (P>0.1). ATRs were stratified into clinical categories of mild (pruritus/urticaria, n=3), moderate (angioedema/dyspnea, n=5), and severe (any symptom with hypotension, n=2). There were no differences or trends in HR and LTC4 production among the categories (P>0.2). There was no correlation between pre/post transfusion tryptase elevations in recipient plasma (n=4) and HR or LTC4 production. HR and LTC4 production was similar among blood types of AP products (A: n=12, O: n=5, AB: n=3), (P>0.7). We previously reported that concentrations of C5a are higher in AP products associated with ATRs than controls. C5a content in AP supernatant modestly correlated with basophil HR release (rho: 0.47, P=0.04), but less with LTC4 production (rho: 0.27, P=0.3). Summary: An in vitro basophil activation test that measures HR and LTC4 production was developed to study ATRs. We find that 1) IL-3 priming of basophils was necessary to elicit responses to AP supernatants; 2) LTC4 and HR are correlated within each reaction; 3) there are no differences in HR and LTC4 production between ATR and control AP supernatants were observed, regardless of clinical ATR severity; and 4) HR is associated with C5a content in the AP supernatant. Conclusions: The observations of 1) an overall requirement for basophil priming and 2) the lack of correlation between the clinical severity of ATRs and basophil HR and LTC4 release implicate recipient atopic priming as a risk factor for ATRs. However, the association of AP C5a content with HR suggests a donor/product role in ATRs, as well. Studies focusing on the nature of recipient susceptibility may further elucidate mechanisms of ATRs. Disclosures: No relevant conflicts of interest to declare.

CD203c expression–based basophil activation test for diagnosis of wheat-dependent exercise-induced anaphylaxis

2012 ◽  
Vol 129 (5) ◽  
pp. 1404-1406 ◽  
Author(s):  
Yuko Chinuki ◽  
Sakae Kaneko ◽  
Itaru Dekio ◽  
Hitoshi Takahashi ◽  
Reiko Tokuda ◽  
...  
Keyword(s):  
Basophil Activation Test ◽  
Basophil Activation ◽  
Activation Test ◽  
Exercise Induced

Usefulness of basophil activation test for the diagnosis of peanut allergy in Japanese children

2014 ◽  
Vol 28 (2) ◽  
pp. 216-225
Author(s):  
Shigeyuki Narabayashi ◽  
Takaaki Meguro ◽  
Yasunori Ito ◽  
Fumika Tokunaga ◽  
Shiro Seto ◽  
...  
Keyword(s):  
Peanut Allergy ◽  
Basophil Activation Test ◽  
Basophil Activation ◽  
Japanese Children ◽  
Activation Test

scholarly journals Innovative robust basophil activation test using a novel gating strategy reliably diagnosing allergy with full automation

Allergy ◽  
2021 ◽  
Author(s):  
Jochen Behrends ◽  
Christian Schwager ◽  
Martina Hein ◽  
Thomas Scholzen ◽  
Skadi Kull ◽  
...  
Keyword(s):  
Basophil Activation Test ◽  
Basophil Activation ◽  
Activation Test ◽  
Full Automation

The basophil activation test identified carminic acid as an allergen inducing anaphylaxis

2013 ◽  
Vol 1 (2) ◽  
pp. 197-199 ◽  
Author(s):  
Naoya Sugimoto ◽  
Masao Yamaguchi ◽  
Yusuke Tanaka ◽  
Yuko Nakase ◽  
Hiroyuki Nagase ◽  
...  
Keyword(s):  
Basophil Activation Test ◽  
Basophil Activation ◽  
Carminic Acid ◽  
Activation Test
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