Lisa Strasser

Abstract

As antigen presenting cells, especially dendritic cells (DCs), are crucial players in the initiation phase of an allergic immune response. Therefore, we analyze the proteome of monocyte derived dendritic cells (moDCs) after stimulation with birch pollen extract (betex) and a recombinant variant of the major birch pollen allergen Bet v 1 (rBet). Thus, we try to elucidate, whether an allergic immune response is triggered by the allergen itself or if the combination of stimulants contained in the pollen (e.g. endotoxins) is required.
To address this question we perform quantitative proteome analysis using iTRAQ®-labeling for relative quantification. Hence, we generate moDCs and stimulate them with betex [10.0 µg/ml], rBet [10 µg/ml] and lipopolysaccharide [1.0 ng/ml]. Subsequently, proteins are extracted and tryptically digested. To reduce donor variability, peptides of several donors are merged prior to the labeling. Samples are then analyzed using reversed-phase (RP) high-performance liquid chromatography (HPLC) and tandem mass spectrometry (MS/MS).