Time-dependent accumulation of prion infectivity in isolated splenic cell types.

‡Myeloid cells were isolated with CD11b magnetic beads after partial depletion of DCs and contain CD11c− CD11b+ macrophages and CD11c+ CD11b+ mDCs (see Figure 1B).†CD11b+ macrophages devoid of mDCs contaminants were isolated by FACS (see Figure 1C).#Infectious titers are represented as mean values ± SE, and as lower and upper limits of 95% confidence intervals (conf.int).Groups of ten 129/Sv×C57BL/6 mice, inoculated i.p. with 100 µl aliquots of 1% (w/v) RML brain homogenate I6200 were culled at various time points after inoculation as specified above and splenocytes and splenic cell types were serially isolated by MACS after Collagenase digestion and Lympholyte purification according to Figure 1. Infectious titers were determined by SCEPA using a GLM as specified above. Mean values ± SE and 95% confidence intervals for at least three independent experiments are shown at 14 and 30 dpi. Data from a single experiment are shown where no SE is reported. The detection limit of the assay for splenic cell types was 0.15 TCIU/Mio, for MACS-isolated cells from whole blood 5 TCIU/Mio.