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Basophil activation test: Mechanisms and considerations for use in clinical trials and clinical practice

Santos, Alexandra F., Alpan, Oral, Hoffmann, Hans‐Jürgen·2021·Allergy
DOI 10.1111/all.14747

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Abstract

The basophil activation test (BAT) is a functional assay that measures the degree of degranulation following stimulation with allergen or controls by flow cytometry. It correlates directly with histamine release. From the dose‐response curve resulting from BAT in allergic patients, basophil reactivity (%CD63 + basophils) and basophil sensitivity (EC 50 or similar) are the main outcomes of the test. BAT takes into account all characteristics of IgE and allergen and thus can be more specific than sensitization tests in the diagnosis of allergic disease. BAT reduces the need for in vivo procedures, such as intradermal tests and allergen challenges, which can cause allergic reactions of unpredictable severity. As it closely reflects the patients' phenotype in most cases, it may be used to support the diagnosis of food, venom and drug allergies and chronic urticaria, to monitor the natural resolution of food allergies and to predict and monitor clinical the response to immunomodulatory treatments, such as allergen‐specific immunotherapy and biologicals. Clinical application of BAT requires analytical validation, clinical validation, standardization of procedures and quality assurance to ensure reproducibility and reliability of results. Currently, efforts are ongoing to establish a platform that could be used by laboratories in Europe and in the USA for quality assurance and certification.

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Image 1
CC-BY-NC · From the PDF
FIGURE 1 Historical timeline of the basophil activation test (BAT). EQA, external quality assurance
FIGURE 1 Historical timeline of the basophil activation test (BAT). EQA, external quality assuranceCC-BY-NC · From the PDF
FIGURE 2 Examples of gating strategies for basophils: (a), Basophils were identified as SSClow CD123c + CD193 + cells 45,48 : 1. Lymphocyte -­ monocyte gate on a FSC/SSC plot using a logarithmic scale, 2. Doublet exclusion FSC-­ H vs. FSC-­ A, then SSC-­ H vs. SSC-­ H, 3. Gate on both markers simultaneously CD123 and CD193, 4. CD63 negative threshold was set to 2.5% and the positive population above that threshold was assessed. (b), Basophils were identified as Lymphocyte/monocyte gate, SSClow CD203c + CD123 + HLA- ­DR- ­. 2,6,26,37 The CD63 gate is set on the negative control and basophil activation is measured above this gate for the other stimulation conditions, either with allergen or positive controls
FIGURE 2 Examples of gating strategies for basophils: (a), Basophils were identified as SSClow CD123c + CD193 + cells 45,48 : 1. Lymphocyte -­ monocyte gate on a FSC/SSC plot using a logarithmic scale, 2. Doublet exclusion FSC-­ H vs. FSC-­ A, then SSC-­ H vs. SSC-­ H, 3. Gate on both markers simultaneously CD123 and CD193, 4. CD63 negative threshold was set to 2.5% and the positive population above that threshold was assessed. (b), Basophils were identified as Lymphocyte/monocyte gate, SSClow CD203c + CD123 + HLA- ­DR- ­. 2,6,26,37 The CD63 gate is set on the negative control and basophil activation is measured above this gate for the other stimulation conditions, either with allergen or positive controlsCC-BY-NC · From the PDF
FIGURE 3 A typical dose-­ response curve of basophil activation with increasing concentrations of allergen of a grass pollen allergic patient from a published study 45,48 is represented. The maximal basophil activation expressed is a measure of basophil reactivity. EC 50 is the concentration eliciting the half-­ maximal basophil activation and is a measure of basophil sensitivity
FIGURE 3 A typical dose-­ response curve of basophil activation with increasing concentrations of allergen of a grass pollen allergic patient from a published study 45,48 is represented. The maximal basophil activation expressed is a measure of basophil reactivity. EC 50 is the concentration eliciting the half-­ maximal basophil activation and is a measure of basophil sensitivityCC-BY-NC · From the PDF
Image 5
CC-BY-NC · From the PDF
FIGURE 5 Proposed sequence of tests to support the diagnosis of food allergy, before referring patient for an oral food challenge. CRD, component-­ resolved diagnosis; BAT, basophil activation test; MAT, mast cell activation test
FIGURE 5 Proposed sequence of tests to support the diagnosis of food allergy, before referring patient for an oral food challenge. CRD, component-­ resolved diagnosis; BAT, basophil activation test; MAT, mast cell activation testCC-BY-NC · From the PDF
Image 7
CC-BY-NC · From the PDF

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Metadata

Authors
Santos, A. F., Alpan, O., & Hoffmann, H.
Affiliations
  • Department of Women and Children's Health (Pediatric Allergy) School of Life Course Sciences Faculty of Life Sciences and Medicine King's College London London UK
  • Amerimmune Fairfax VA USA
  • Department of Clinical Medicine Aarhus University Aarhus Denmark
ORCID
Venue
Allergy
Publisher
Wiley
Published
2021-02-27
Volume
76
Issue
8
Pages
2420-2432
ISSN
  • 0105-4538
  • 1398-9995
DOI
10.1111/all.14747
Cited by
273
References
4
Language
en
Licence
creativecommons.org/licenses/by-nc/4.0/ (vor)

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