Figure 3. Clone 13 efficiently infects CD8α^neg CD11c^posDC.

Adult C57BL/6 were infected with LCMV Arm, or Cl13 and spleens harvested 7 days post infection. CD11c^pos cells were positively selected by MACS bead isolation following CD19/90 depletion. (A) Isolated cells were stained for CD11c, CD90, CD19, CD8α and cell surface (top two rows) LCMV NP or intracellular (bottom row) LCMV NP. Gating controls are as follows: Naïve DC (dashed line), DCs from Cl13 infected mice incubated with isotype control antibody for LCMV NP Ab (filled grey histogram). (B & C) CD8α^neg DCs were sorted based on cell surface NP expression. (B) RNA was extracted from sorted DCs; reverse transcription was performed and generated cDNA used in Real Time PCR reaction to determine the copy number of LCMV GP, relative to known concentration of control plasmid. ND indicates below detection limits (C) 4×10⁴ sorted DCs were cultured for 2 days in cRPMI. IL-10 levels were measured in culture supernatants of Cl13 infected (NP^pos), exposed (NP^neg) DCs and Naïve CD8α^neg DCs. n = 4–5 mice per treatment per experiment. RealTime and ELISA data from one representative of three independent experiments is shown. Standard deviation is shown.