Anaphylaxis is a severe, potentially life-threatening allergic reaction, which can occur within seconds or minutes of exposure to an allergen. Mast cells and basophils are believed to be the main effector cells of anaphylaxis. Both cells express high affinity IgE receptor (FcεRI) on their surface. Recent data suggest a substantial migration of circulating basophils during anaphylaxis, which correlates with a significant increase in serum concentrations of the chemokine CCL2. The aim of our study was to determine the effect of serum anaphylactic factors and CCL2 on migration of basophils in vitro. Basophils were isolated with negative immunomagnetic selection from peripheral whole blood of healthy donors. Paired serum samples of patients taken during and after anaphylaxis were used. Migration of basophils took place in adapted Boyden chambers at 37 °C and 5 % CO2. Anti-CCL2 neutralizing antibody was used to block CCL2. Analysis of the number and activation of basophils was carried out on a flow cytometer. Our results showed that there was a significant difference in the basophil activation in heparinized blood compared to EDTA blood. We showed that serum anaphylactic factors have impact on basophil migration, but we could not confirm that the blockade of CCL2 in serum results in reduced migration. We also confirmed that human recombinant CCL2 (rCCL2) in buffer solution promotes basophil migration, whereas anti-CCL2 treatment reduces it. Further studies are needed to elucidate the exact role and source of CCL2 during anaphylaxis.
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