Global gene expression profiles of basophils isolated from human blood after engagement of immunoglobulins on their surface.

Basophils are blood leukocytes associated to Th2 responses and allergy reactions primarily through the IgE bound to their surface throught the Fc Epsilon hifh affinity receptor. Soluble IgD is an enigmatic antibody isotype whose function remains elusive. We have characterised the presence of this antibody isotype on the surface of basophils where we hypothesise that it plays a role in Th2 responses and allergy reactions. Basophils from peripheral blood from 3 donors were MACS sorted (Basophil isolation Kit II, Milteny) and to increase purity we performed negative FACSorting using lineage-specific antibodies (CD3, CD14, CD19, CD56) (Final purity >98%). These were either untreated or treated with anti-IgE, anti-IgD or both, for 20 minutes at 37º before proceeding to RNA extraction.