Activation markers for human basophils

Section: Methodsmentioning

confidence: 99%

Differential modulation of human basophil functions through prostaglandin D₂ receptors DP and chemoattractant receptor‐homologous molecule expressed on Th2 cells/DP2

Yoshimura-Uchiyama

Iikura²,

Yamaguchi³

et al. 2004

“…basophils, mast cells, macrophages and platelets [26, 27]. In resting basophils, CD63 is anchored in basophilic intracytoplasmic granules and is generally weakly expressed on the surface membrane both in normal subjects as well as in patients with allergies [28–31]. In contrast, as a result of fusion between the granule and plasma membrane, CD63 is expressed with a high density on activated basophils and mirrors histamine release [23, 32, 33].…”

Section: Flow Cytometric Analysis Of In Vitro‐activated Basophils: Prmentioning

confidence: 99%

“…In addition, priming with IL‐3 has been observed to increase assay sensitivity [43, 44]. Although in recent studies priming with IL‐3 alone did cause little or no histamine release [45–48] or CD63 up‐regulation [30], non‐specific basophil activation leading to false‐positive results needs to be kept in mind. This area actually requires further comparative investigation.…”

Section: Flow Cytometric Analysis Of In Vitro‐activated Basophils: Tementioning

confidence: 99%

In vitro allergy diagnosis: should we follow the flow?

Ebo

Hagendorens

Bridts

et al. 2004

113

During the last 5 years, an increasing number of studies have demonstrated that flow cytometric quantification of in vitro basophil activation can be a quite performant and reliable tool to measure IgE-dependent allergen-specific responses in allergic patients. So far, most assays have used CD63 as a basophil activation marker and native allergen extracts for stimulation. However, other basophil markers and recombinant allergens have recently been introduced. The technique has been applied for the diagnosis of allergy to pollen, house dust mite, food, natural rubber latex, hymenoptera venom and drugs. In addition, the technique has proven to be useful in non-IgE-mediated reactions such as hypersensitivity to drugs as well as detection of auto-antibodies in chronic urticaria. This review will focus on some specific issues: (1) principles of flow cytometric analysis of in vitro-activated basophils, (2) general technical aspects of the technique (including passive sensitization), (3) clinical applications and (4) recommendations for further development and evaluation of the technique.

“…Unlike CD63, its expression is unique to basophils, mast cells and their progenitors [16, 17]. CD69 is another surface marker that is increased on basophils following IgE‐mediated activation [28]. However, because the expression of this molecule is also increased following IL‐3 exposure, its use is limited in determining IgE‐dependent reactions.…”

Section: Introductionmentioning

confidence: 99%

Effect of in vitro aspirin stimulation on basophils in patients with aspirin‐exacerbated respiratory disease

Çelik

Schroeder

Hamilton

et al. 2009

Background Basophil activation has been implicated in the pathogenesis of aspirin exacerbated respiratory disease. However, a comprehensive analysis of basophil responses to aspirin in terms of mediator release, cytokine secretion and increased expression of surface activation markers has not been performed. Objective To study the in vitro effects of aspirin on the concurrent release of histamine, leukotriene C4 and IL-4 from human basophils and to also evaluate changes in surface activation markers (CD63, CD69 and CD203c) expressed by these cells. Methods Basophil-enriched cell suspensions from 10 patients with aspirin exacerbated respiratory disease and 10 healthy volunteers were incubated with lysine-aspirin for up to 3 hours. Cells were analysed for expression of CD63, CD69 and CD203c using flow cytometry. Cell-free supernatants were evaluated for histamine, and leukotriene C4 release and for IL-4 secretion. Results Aspirin-induced expression of CD63, CD69 and CD203c yielded 30%, 80% and 70% sensitivity, respectively, but with poor specificity. There was no significant difference in leukotriene C4 synthesis between groups. None of the patients with aspirin exacerbated respiratory disease (or controls) released IL-4 in response to aspirin. A higher dose of 5 mg/ml aspirin mediated non-specific effects on basophils. Conclusion Basophil responses to in vitro aspirin challenge are poor indicators of clinical sensitivity. Aspirin activates some basophils by means of mechanisms which differ from the classical IgE mediated pathway. Our study also shows that the use of 27 mM of aspirin (5 mg/ml) used by previous investigators causes nonspecific basophil activation, thereby eliminating its usefulness in a cell based diagnostic test for AERD. Evaluation of in vitro basophil activation has low clinical value in identifying aspirin- induced respiratory reactions