Major pitfalls in BAT performance may be caused by gating protocols and CD63% cut off evaluation

@article{Chirumbolo2014MajorPI,
  title={Major pitfalls in BAT performance may be caused by gating protocols and CD63\% cut off evaluation},
  author={Salvatore Chirumbolo},
  journal={Cytometry Part A},
  year={2014},
  volume={85}
}
A paper by Ariza et al., published in the current issue of Cytometry Part A, described the ability of basophil activation test (BAT) in diagnosing patients with cross-intolerance to NSAIDs (1). Ariza et al. concluded that BAT used in their study did not appear useful to evaluate patients with NSAIDderived allergy, despite the fact that their flow cytometry approach to investigate basophils by a CD193/CD203c/CD63 BAT represented a novelty in the field. The data reported by the authors (1… 
BAT in the Anti-histamine Drug Allergy
TLDR
A phenotyping protocol to capture basophils in the blood sample of the pregnant woman and a CD63 pos marker as the activation probe and a SSC low /CD203c pos gating strategy is used.
Is basophil activation test (BAT) really useful for allergy diagnosis?
  • S. Chirumbolo
  • Medicine, Biology
    European annals of allergy and clinical immunology
  • 2016
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It is practically impossible to set basophil activation on a standard resting population, because of the many reasons related to pre-analytical handling, frequency of releaser or non releaser subjects within the population, intrinsic variability due to CD63 vesicle endowment.
Basophil activation test: handle with care.
  • G. Maietta
  • Biology, Medicine
    European annals of allergy and clinical immunology
  • 2016
TLDR
Basophil activation test should be considered as an additional clinical tool in the diagnosis of allergy and not the “absolute truth”, as highlighted in the letter to the Editor by Dr Chirumbolo.
Using a CD45dim/CD123bright/HLA-DRneg phenotyping protocol to gate basophils in FC for airway allergy. CD123 does not decrease.
TLDR
The methodological aspects of the use of the protocol adopting the panel CD45dim/CD123bright/HLA-DRneg to gate basophils in flow cytometry are described, trying to highlight the main biases related to an incorrect use of this protocol.
Commentary: The Expression of CD123 Can Decrease with Basophil Activation: Implications for the Gating Strategy of the Basophil Activation Test
TLDR
It is shown that, during cellular activation, the expression of membrane CD123, the alpha subunit of the interleukin-3 (IL-3) receptor, is downregulated, leading to a possible loss in the electronic capture of basophils and hence a false negative outcome of the activation test.
On the reliability of the CD123‐endowed basophil activation test (BAT) and its application in food allergy
TLDR
It is concluded that BAT should be preferentially used in association with high protein sesame extract-skin prick test (HSPE-SPT) for the diagnosis of sesame food allergy, so preventing any ethical issue regarding the possble risk associated with oral food challenge (OFC).
In Vitro Diagnosis of Immediate Drug Hypersensitivity Anno 2017: Potentials and Limitations
TLDR
Although drug-sIgE assays and BAT can provide useful information in the diagnosis of IDHR, their predictive value is not absolute.
The role of basophil activation test in allergic bronchopulmonary aspergillosis and Aspergillus fumigatus sensitization in cystic fibrosis patients.
Basophil activation test in oral desensitization to cow’s milk allergy
  • S. Chirumbolo
  • Medicine
    United European gastroenterology journal
  • 2016
TLDR
Two standpoints in the CD63 response to food allergy and OAT and their large difference inCD63 response before and after suggests for the optimal performance of a CD123/HLADR/CD63 BAT in oral food allergy immunotherapy.
Reply to Claude Lambert
TLDR
A debate in this issue of Human Vaccines and Immunotherapeutics about basophil activation test (BAT) in immunotherapy provided the opportunity to shed a light on issues that may raise further criticisms or any misunderstanding from readers of the Journal.
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TLDR
There is now accumulating evidence indicating that the basophil activation test (BAT), using either CD203c or CD63 as activation markers, has become a robust and reliable tool usable in clinical practice for the in vitro investigation of immediate allergy.
The use of IL‐3 in basophil activation tests is the real pitfall
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The article tries to assess the use of basophil activation tests (BATs) based on CD203c upregulation as a reliable tool for allergy diagnosis and suggests how to use BATs that evaluate CD203C upregulation.
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Comparison of CD63 Upregulation Induced by NSAIDs on Basophils and Monocytes in Patients with NSAID Hypersensitivity
TLDR
Positive tests were more frequent among patients having a severe hypersensitivity contrasting with the other patients who had results similar to controls, and with aspirin, the sensitivity and specificity of BAT agree with already published results.
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TLDR
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TLDR
Use of polychromatic flow cytometry allowed efficient basophil electronic purification and identification of different behaviors of the major activation markers, including CD203c as a more sensitive marker than CD63, in response to fMLP but not inresponse to anti-IgE and to calcium ionophore.
Threshold for basophil activation test positivity in neuromuscular blocking agents hypersensitivity reactions
TLDR
With a stimulation index ≥ 1.76 and a percentage of activated basophils > 5.01% as threshold, the performance of BAT for NMBAs yields 68.18% sensitivity and 100% specificity.
Diagnostic Tests Based on Human Basophils: Potentials, Pitfalls and Perspectives
TLDR
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TLDR
Overall the data show that CCR3 demonstrates adequate sensitivity but weak specificity in its ability to reliably identify histamine-releasing activated basophils.
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