Figure 4.

C3a and C5a are present in plasma-derived FVIII products. (A) Plasma-derived (pd) or recombinant (r)FVIII products (5 IU each) were analyzed for C3a (black bars) and C5a (white bars) by enzymelinked immunosorabant assay (ELISA) with water for injection (WFI) as solvent control. Error bars indicate standard deviations from triplicate or quadruplicate ELISA measurements. (B) Dendritic cells (DC) were treated for 24 hours with rFVIII (1 IU/mL), lipopolysaccharide (LPS) (0.1 mg/mL), or with rFVIII plus LPS in the presence or absence of plasma or heat-treated plasma (both at 2.5 mL/mL). Control DC were left untreated. PKH26-labeled autologous T cells were added and 9 days after co-culture, cells were harvested and percentages of proliferated T cells were analyzed by fluorescence-activated cell sorting. Data of 1 representative donor are shown. (C) Summarized results of several donors from 2 independent experiments are given (n=4). Error bars indicate the means ± standard error of the mean. SSC: side scatter; FSC: forward scatter.