Stimulation of TLR2 and TLR4 differentially skews the balance of T cells in a mouse model of arthritis
J. Clin. Invest. Shahla Abdollahi-Roodsaz, et al. 118:205 doi:10.1172/JCI32639 [Go to this article.]

Figure 4
Influence of Tlr2 deficiency on various Th cell subsets and the expression of related cytokines. mRNA expression of T cell markers in inflamed synovium of the ankle joints (A) and spleens (B) of IL1rn^–/–Tlr2^–/– compared with IL1rn^–/–Tlr2^+/+ mice. Synovial biopsies were pooled from more than 5 joints of 15-week-old mice, and spleens were from 4 nonarthritic 6-week-old mice. mRNA expression was measured by quantitative PCR. Relative mRNA expression compared with the housekeeping gene GAPDH (2^-dCt × 1,000) is shown on the y axis. IFN-γ (C) and IL-17 (D) production by spleen and lymph node cells upon 72 hours stimulation with anti-CD3 (0.5 μg/0.2 ml/well) and anti-CD28 (2 μg/ml), measured by Luminex; results are mean ± SEM from a representative experiment with n > 4 mice per group. Protein expression of Foxp3 (E) and CD25 (F) on CD4⁺-gated CD25⁺Foxp3⁺ cells from whole blood and spleen of 15-week-old mice after correction for isotype-matched IgG control; mean ± SEM of n = 3 from a representative experiment. (G and H) T cell suppression assay on splenic cells of IL1rn^–/–Tlr2^+/+ and IL1rn^–/–Tlr2^–/– mice. (G) CD4⁺CD25^– Teffs (50,000 cells/well) were stimulated with 1 μg/ml anti-CD3, 2 μg/ml concanavalin A, or 60 IU/ml IL-6 (negative control) in the presence of 50,000/well irradiated CD4^– cells as APCs. After 3 days, proliferation was measured by [³H]thymidine incorporation. (H) For suppression assays, cells were incubated with anti-CD3 and APCs in the presence of titrated numbers of Tregs (CD4⁺CD25⁺). Percent suppression was calculated relative to the cultures without Tregs. Measurements were performed in triplicate, and values shown are mean ± SEM of 4 mice per group. mRNA expression of TGF-β1 in noninflamed and inflamed ankle synovium (I) and spleen (J), measured by quantitative PCR. Relative mRNA expression compared with GAPDH is shown. *P < 0.05.