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 6
Decreased IL-17 in IL1rn^–/–Tlr4^–/– mice and IL-1–mediated effects of TLR4 activation on IL-23/IL-17 production. (A–D and F) Spleens and lymph nodes from 5- to 6-week-old mice without arthritis were isolated and disrupted. CD3⁺ T lymphocytes were isolated using magnetic beads (MACS). (A and B) Th17 FACS analysis following stimulation with PMA (50 ng/ml), ionomycin (1 μg/ml), and brefeldin (1 μl/ml) for 5 hours ; n = 4. (C) IL-17 production by splenic and lymph node T cells upon stimulation with anti-CD3 (0.5 μg/0.2 ml/well) and anti-CD28 (2 μg/ml) for 72 hours; measured by Luminex; n ≥ 6. (D) TLR4 activation of bone marrow–derived DCs by 100 ng/ml LPS for 24 hours resulted in higher IL-23 production in IL1rn^–/– cells compared with BALB/c WT. IL-23 was measured by ELISA; n = 10. (E) TLR4 (200 ng/ml LPS) plus anti-CD3 (1 μg/ml) activation of splenic T cells leads to higher IL-1–mediated IL-17 production in IL1rn^–/– cells compared with WT cells, as measured by Luminex; n = 5. (F) Bone marrow–derived DCs from IL1rn^–/–Tlr4^–/– mice are not compromised in IL-23 production upon non-TLR4 stimulations. DCs were stimulated with 100 ng/ml LPS, 100 ng/ml Pam3Cys, or a cocktail of IL-1β (25 ng/ml), TNF-α (25 ng/ml), IL-6 (100 ng/ml), and PGE2 (1 μg/ml) for 24 hours; n = 4 per group. Data are mean ± SEM. NRS, normal rabbit serum. *P < 0.05, **P < 0.01, and ***P < 0.001.