Skip to main content
. 2014 Nov 24;97(2):379–390. doi: 10.1189/jlb.4A0714-339R

Figure 6. B. burgdorferi-induced type I and type II IFNs cooperate to drive production of enzymatically active IDO in human PBMCs.

Figure 6.

Human PBMCs were coincubated for 24 h at 37°C with B. burgdorferi strains B31-A3 and A3-M9 lp36− at a MOI of 10:1. Where indicated, B18R, a soluble vaccinia virus-encoded IFN-αR, and α-IFNGR1 were added to PBMCs, 1 h before stimulation with B. burgdorferi. (A) PBMC lysates were resolved by 12.5% SDS-PAGE for Western immunoblotting with rabbit anti-human IDO1. Densitometric values of protein expression were normalized to GAPDH and quantitated by use of ImageJ software. (B) Secreted kynurenine concentrations in cell-free culture supernatants were determined by a spectrophotometric assay and interpolated from a standard curve of known kynurenine concentrations. Columns depict the mean ± sd of results from 1 donor assessed in triplicate. Representative Western blot images are shown. Statistical analysis was performed by use of a one-way ANOVA with Tukey-Kramer’s post-test. ***P < 0.001; *P < 0.05, relative to PBMCs incubated with medium alone.