FIGURE 1.

Freshly-isolated plasmacytoid (p)DC express higher levels of NOD2 message than myeloid (m) DC and in vivo treatment with MDP does not affect the number and proportion of liver and spleen DC subsets. Freshly-isolated, immunobead-purified liver and spleen pDC and mDC were isolated from C57BL/10 (B10) mice treated for 10 consecutive days with the endogenous DC poietin Flt3L (10µg/d, i.p.). A) mPDCA-1⁺ and mPDCA-1⁻CD11c⁺ immunobead-purified cells were stained with anti-B220 and anti-CD11c mAbs. Flow cytometric analysis shows a purity >95% of liver and spleen CD11c^lowB220⁺ pDC and CD11c⁺B220⁻mDC; B) DC subsets were analyzed for NOD2 relative to β-actin mRNA expression by semi-quantitative RT-PCR, as described in the Materials and Methods. Data are representative of three independent experiments, with two mice per group in each experiment. *p<0.004. For negative and positive control purposes, NOD2 expression was evaluated in a mouse D10 T cell line (Tc) and in normal mouse peritoneal macrophages (M), respectively. (C) Absolute number of Flt3L-expanded, freshly-isolated, immunobead-purified liver and spleen pDC and mDC from MDP-treated (100µg/d, i.p., 3 days) and control (PBS) mice. Data are representative of five independent experiments, with two mice per group in each experiment.