Inhibition of dendritic cell-mediated antigen-specific Th1 cell responses by lidocaine <i>in vivo</i>.

<p>Bone marrow-derived dendritic cells were pulsed with OVA_323-339 in the presence of lidocaine or vehicle before being transferred into OT-II TcR transgenic mice (n = 3~4). (A and B) The frequencies of IFN- γ producers among Vα2⁺ cells. (C) The amounts of the indicated cytokines in the supernatant of splenocyte stimulated with OVA_323-339 were measured by ELISA. Data represent two independent experiments. Data shown are mean ± SEM. *<i>p</i><0.05.</p

Lidocaine inhibits dendritic cell-mediated Th1 cell differentiation while having little effects on dendritic cell-mediated Th2, Th17, regulatory T cell differentiation <i>in vitro</i>.

<p>Naïve CD4⁺ T cells were co-cultured with bone marrow-derived dendritic cells with Th1, Th17, Th2 or regulatory T cell differentiation conditioned-media or cultured with plate-coated anti-CD3 and anti-CD28 with supernatant of dendritic cells stimulated with LPS in the presence of lidocaine (0.2 mg/ml or indicated dose) or vehicle. (A-D) The frequencies of IFN-γ, IL–17, IL–4/5 or Foxp3 positive cells among CD4⁺ population were measured by flow cytometer. (E) The level of IFN-γ was measured using co-cultured supernatants from Th1 differentiation condition. Data represent at least two independent experiments. Data shown are mean ± SEM. *<i>p</i><0.05; **<i>p</i><0.01; ***<i>p</i><0.001; NS, not significant.</p

Lidocaine inhibits dendritic cell-mediated Th1 cell differentiation <i>in vitro</i>.

<p>Naïve CD4⁺ T cells were either co-cultured with bone marrow-derived dendritic cells in the presence of soluble anti-CD3 and LPS, or in anti-CD3, CD28 pre-coated plates in the presence of IL–2 and IL–12 for Th1 cell differentiation. Lidocaine was added at a concentration of 0.2 mg/ml. (A & B) The frequencies of IFNγ or IL–17 producing cells among CD4⁺ T cells. (C) The mRNA levels of the indicated genes. (D) The levels of IFN-γ in the cultured supernatants of naïve CD4⁺ T cells cultured with vehicle- or lidocaine-conditioned media. Data represent at least three independent experiments. Data shown are mean ± SEM. **<i>p</i><0.01; ***<i>p</i><0.001; NS, not significant.</p

Effects of lidocaine on the expression of various cytokines upon LPS stimulation.

<p>Bone marrow-derived dendritic cells were stimulated with 100 ng/ml of LPS in the presence of vehicle (EtOH) or 0.2 mg/ml lidocaine for 4 h and 24 h to examine mRNA expression and cytokine production, respectively. (A) The mRNA levels of the indicated genes were analyzed by quantitative RT-PCR. (B) The amounts of each cytokine produced were measured by ELISA. All experiments were performed at least three times. Data shown are mean ± SEM. *<i>p</i><0.05; ***<i>p</i><0.001; ND, not detected.</p

Lidocaine regulates the expression of cytokines and NF-κB signaling pathway in a dose-dependent manner.

<p>(A) Bone marrow-derived dendritic cells were stimulated with 100 ng/ml of LPS together with increasing concentrations of lidocaine for 4 h. The mRNA levels of the indicated genes were analyzed by quantitative RT-PCR. (B) Raw 264.7 cells were treated with increasing doses of lidocaine for 2 h and stimulated with LPS for 20 min. The expression of IκB-α was examined by western blot. All experiments were performed at least three times. Data shown are mean ± SEM. *<i>p</i><0.05; **<i>p</i><0.01.</p

Regulation of cytokines expression in dendritic cells in response to various TLR ligands by lidocaine.

<p>(A & B) Bone marrow-derived dendritic cells were stimulated with LPS (100 ng/ml), poly(I:C) (1 μg/ml) or R837 (1 μg/ml) in the presence of lidocaine (0.4 mg/ml) or vehicle. The amounts of IL–6 and TNF-α in the supernatant were measured by ELISA. (C) The mRNA levels of the indicated genes were analyzed by quantitative RT-PCR. Data represent at least two independent experiments. Data shown are mean ± SEM. *<i>p</i><0.05; **<i>p</i><0.01; ND, not detected.</p