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cea4030-sup-0001-FigS1.pdfapplication/PDF70KFigure S1. Effectiveness of cell separation. PBCMs from allergic patients were depleted of or enriched for CD20, CD27, CD38, CD126 or CD203c using magnetic separation, stained with the respective antibodies and subjected to flow cytometry. Gates indicate the percentage of positive staining for the respective antibodies in total PBMCs (upper panels), depleted (middle panels) or enriched populations (lower panels). Blots are representative for cell preparations from 8 allergic patients.
cea4030-sup-0002-FigS2.pdfapplication/PDF25KFigure S2. Southern blot for IgE. Nitrocellulose-blotted IgE heavy chain-encoding cDNAs were hybridized with a 32P-labelled oligonucleotide probe specific for Cε1. Lanes are as follows: (A) 1: Total PBMCs, 2 and 3: CD138-depleted sample 1 and 2, 4: CD138-enriched, M: DNA ladder mix.
cea4030-sup-0003-TableS1.pdfapplication/PDF9KTable S1. IgE is stable for 1 week in supernatants of PBMC cultures. PBMCs from three patients (ID9-11) were cultured in medium containing 20% serum from allergic patients. Total IgE levels were measured by ImmunoCAP (kUA/L) in PBMC supernatants (15×106 cells/3 ml) on day 0 and day 7 of culture.
cea4030-sup-0004-TableS2.pptapplication/70KTable S2. Allergen-specific IgE levels in supernatants of allergen-stimulated PBMC cultures from 5 allergic patients. PBMC cultures were either left unstimulated or stimulated for 1 week with two different allergens (250 ng/mL) to which the patient was sensitized to. Allergen-specific IgE levels were measured by ultrasensitive ImmunoCAP measurements (kUA/L) in PBMC supernatants (15×106 cells/3 ml) on day 7 of culture.

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