Host irradiation is prerequisite for pDC maturation to prime T cells. (A) Expression of CD86 and I-Ab expression on B cells, pDCs, and cDCs in the spleen and mLNs isolated from irradiated mice 10 hours after irradiation (filled histograms) and from unirradiated control mice (broken-lined open histograms). (B) Unirradiated (top) or irradiated (bottom) B6-Ly5a (CD45.1+) mice were injected with 80 × 106 BM cells and splenocytes isolated from FL-treated B6 (CD45.2+) mice. Expression of I-Ab on CD45.2+ B cells and pDCs in mLNs 10 hours after injection is shown. (C) Aliquots of 20 × 106 CFSE-labeled BALB/c (H-2Kd+) CD4+ T cells were transferred to unirradiated H2-Ab1−/− mice preinjected with 6 × 106 WT pDCs or cDCs. Cell division of donor CD4+ T cells in the spleens on day +6 is shown. (D) Similarly, CFSE-labeled BALB/c CD4+ T cells were transferred to unirradiated H2-Ab1−/− mice preinjected with freshly isolated pDCs or pDCs stimulated with CpG 1668 1 μM for 24 hours in vitro. Cell division of donor CD4+ T cells in the spleens on day +6 is shown. Data are representative of 2 similar experiments.