Figure 6
Bone marrow (BM)-derived dendritic cells (DCs) from C5aR−/− mice induce decreased proliferation and survival of DO11.10 Rag2−/− T cells in vivo. (a) Experimental design. CD4+ T cells were purified from the spleens of DO11.10 Rag2−/− mice. In all, 5 × 106 T cells were labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) and intravenously (IV) injected into wild-type (wt) BALB/c recipient mice. Twenty-four hours later, mice were intratracheally treated with 106 wt or C5aR−/− BMDCs pulsed with 1 μM DQ-OVA (ovalbumin) for 24 h. Five or nine days later, single-cell suspensions from the lungs and lymph nodes were prepared and the proliferation and the frequency DO11.10 TCR+ (T-cell receptor) T cells was determined. (b) DO11.10 TCR+ T cells in the lymph nodes on day 5 (first upper panel on the left) or day 9 (first lower panel on the left) after adoptive transfer of either wt or C5aR−/− BMDCs; overlay showing the CFSE signal within the DO11.10 TCR+ T cells on day 5 (first upper panel on the right) or day 9 (first lower panel on the right) after adoptive transfer of wt (red) or C5aR−/− (blue) BMDCs. Histograms are representative of ⩾3 independent experiments. (c) Frequency of DO11.10 TCR+ T cells in the lymph node on day 5 (upper panel) or day 9 (lower panel) after adoptive transfer of wt or C5aR−/− BMDCs. (d) DO11.10 TCR+ T cells in the lung on days 5 or 9 after adoptive transfer of wt or C5aR−/− BMDCs. Histograms are representative of ⩾3 independent experiments. (e) Frequency of DO11.10 TCR+ T cells in the lung on days 5 or 9 after adoptive transfer of wt or C5aR−/− BMDCs. Values shown are the mean±s.e.m.; n⩾3 per group, ***P<0.001. GM-CSF, granulocyte macrophages colony-stimulating factor; SSC, side scatter. The color reproduction of this figure is available on the Mucosal Immunology online.
