Figure 6.

Basophil activation in whole blood from donors with confirmed specific IgE-sensitized immune cells against house dust mite (HDM) extract. Whole blood cells were pre-incubated with 0.5 or 5 µM α-13’-COOH or buffer (untreated control) for 50 min. Blood cells used as controls were pre-incubated with buffer. A second unspecific basophil activation was tested either with fMLP (5 µM), C5a (10 nM), or buffer. Activation of basophils with HDM extract was investigated as specific IgE-mediated response. The HDM extract was applied in two concentrations (HDM low: 2 ng; HDM high: 20 ng). A specific anti-IgE receptor mAb was used as positive control (mAb). Activated basophils were stained with a mixture of monoclonal antibodies against human cluster of differentiation (CD) 63 labeled with fluorescein isothiocyanate (anti-CD63-FITC) and against human C-C chemokine receptor type 3 (CCR3) labeled with phycoerythrin (anti-CCR3-PE). Basophil activation is given as the event of activated CD63-positive basophils in relation to the absolute events of each sample in percent. All data are presented as mean ± SEM. Two independent experiments were performed as replicates. Asterisks [*] indicate significant deviations from the untreated control (*** p < 0.001).