Figure 2.

Higher ratios of host plasmacytoid to conventional DCs remain and conventional DCs have a higher expression of CD80 and CD86 after BEN+TBI compared to CY+TBI conditioning. BALB/c mice received 40 mg/kg BEN i.v. or 200 mg/kg CY i.p. day −2 and 400 cGy TBI on day −1. Some groups were transplanted on day 0 with 10⁷ BM + 3 × 10⁶ total T-cells for post-transplant analysis of DCs. On day 0, +1 or +3, spleens were collected and DCs were isolated by MACS negative selection and counted before analysis by flow cytometry. Naïve mice were used as controls. (A) Representative flow cytometry gating of conventional (CD11c+B220–) and plasmacytoid (CD11c+B220+) DC subsets are shown. Of total host CD11c+ DCs, (B) percent pDCs and (C) percent cDCs were quantified on day 0, +1 and +3. Representative histograms, percent and MFI among cDCs positively expressing the activation markers (D) CD80, (E) CD86 and (F) CCR7 are shown, with fluorescence minus one (FMO) control shown in gray, BEN+TBI shown in blue and CY+TBI shown in red. Pooled data from 2 experiments are shown, n = 4–6 mice per group per time point. Mann-Whitney unpaired t-tests were used to determine significance with SEM shown. *P < 0.05, **P < 0.01, ****P < 0.0001.