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. Author manuscript; available in PMC: 2012 Oct 8.
Published in final edited form as: J Immunol. 2006 Nov 1;177(9):5956–5967. doi: 10.4049/jimmunol.177.9.5956

Figure 7. Inoculated Bid KO cDCs promote augmented T cell activation in vivo.

Figure 7

Bid KO and WT mouse ears were tape-stripped and painted with 25μg OVA or HEL in PBS. After an hour, mice were sacrificed and ear specimens harvested for explant culture. Migratory cDCs were cultured from specimens over 48 hours in the presence of OVA (100μg/ml) or HEL (100μg/ml). One million CFSE-labeled CD4 OT-2 T cells were injected via tail vein to WT mice (In this experiment, 50% of the CFSE labeled CD4 cells were OT-2 TCR negative, as indicated by absence of TCR Vβ5 staining - and served as an internal specificity control) and cDCs (5 × 104) were inoculated into the footpad the same day. Popliteal lymph nodes were harvested from individual mice (3 per group) and weighed before processing for TCR and CD69 staining. Flow cytometric analysis plots of (A) the gate used for CFSE-TCR Vβ5 cells, (B) the level of CFSE dilution (indicating cell divisions) and (C) the level of T cell activation/maturation-indicated by transient CD69 up then down regulation are shown. (D) Significant differences in the rate of proliferation between WT and Bid KO. Percent of OT-2 cells found in each division from 3 mice per group is shown (open bar WT, closed bar Bid KO). Students’ T-test P values are depicted as follows: *, P < 0.05; **, P<0.001; n.s., not significant. CFSE T cells showed no proliferation or activation in mice receiving HEL-pulsed LCs and represented 0.01% of total lymph node cells (data not shown). OT-2 T cells expanded from 0.55% to 0.73% of total lymph node cells in mice receiving OVA-pulsed cDCs from WT versus Bid KO mice, respectively (shown in panel A). P<0.05, n=3.