Brucella Control of Dendritic Cell Maturation Is Dependent on the TIR-Containing Protein Btp1 (original) (raw)
Figure 2
Survival and Replication of B. abortus in DCs
(A) BMDCs infected with wild-type B. abortus, _cgs_− mutant, _virB9_− mutant or the wild-type S. typhimurium were lysed and intracellular CFUs enumerated at different times after inoculation. Graph on the right corresponds to bacterial CFU counts performed in CD11c-positive cells infected with either wild-type B. abortus or _virB9_− mutant. Statistical significance was obtained at 24 and 48 h between wild-type and the virB mutant (p < 0.002) and also between wild-type Brucella and Salmonella at 24 h (p = 0.047) and 48 h (p < 0.002).
(B) Quantification of the percentage of wild-type or _virB9_− mutant BCVs that contain LAMP1 by confocal immunofluorescence microscopy. The difference between wild-type and the virB mutant were always statistically significant (p = 0.045 at 30 min and 4 h; p < 0.002 for other timepoints).
(C) Representative confocal images of DCs infected for 24 h with wild-type B. abortus or _virB9_− mutant expressing GFP, labelled with antibodies against MHC II (blue) and LAMP1 (red).
(D) Confocal image of DCs infected for 24 h with wild-type B. abortus expressing GFP, labelled with antibodies against MHC II (blue) and KDEL (red). Samples were also processed for conventional electron microscopy (E), cytochemistry for glucose 6 phosphatase detection (F) or immunogold labelling with an anti-calnexin antibody (G). Bars: 5 μm (C and D); 0.5 μm (F and G). For (A) and (B) data are means ± standard errors of four independent experiments.