Brucella Control of Dendritic Cell Maturation Is Dependent on the TIR-Containing Protein Btp1 (original) (raw)
Figure 7
B. abortus Protein Btp1 Modulates TLR Signaling
(A) Representative images obtained by immunofluorescence microscopy of DCs from wild-type mice or TLR2 knockout mice labelled for MHC II (blue), DALIS (red), and in the case of lower panel also labelled for Brucella (green). Cell were either stimulated with PAM (500 ng/μl, upper panel) or infected with the Brucella wild-type and _btp1_− mutant strains (lower panel).
(B) DCs obtained from wild-type mice or different knockout mice were infected for 24 h with either B. abortus wild-type or _btp1_− mutant strain. The percentage of DALIS in infected cells was then quantified and the data represent means ± standard errors of four independent experiments. The average value for the wild-type Brucella is indicated with a red line. For _btp1_− we observed a statistical difference (*) between the wt and TLR2−/− DCs (p = 0.031).
(C) HEK293 cells were transiently transfected for 24 h with three vectors: a luciferase reporter vector and either empty vector, myc-Btp1 or myc-PipB2 along with either TLR2 (upper panel) or TLR9 (lower panel). The amounts of Btp plasmids are indicated in the graph (ng). Data represent the means ± standard errors of relative luciferase activity obtained from triplicates of a representative experiment. In the case of TLR2, cells were stimulated with PAM (500 ng/μl) and for TLR9 with CpG (1 μM) for 6 h before measuring the luciferase activity.