Department of Biology, Concordia University, Montréal, QC, Canada.
Department of Health, Kinesiology and Applied Physiology, Concordia University, Montréal QC, Canada.
Front Immunol. 2024 Oct 9;15:1446424. doi: 10.3389/fimmu.2024.1446424. eCollection 2024.
Adrenergic receptors regulate metabolic, cardiovascular, and immunological functions in response to the sympathetic nervous system. The effect of β-adrenergic receptor (AR) as a high expression receptor on different subpopulations of T cells is complex and varies depending on the type of ligand and context. While traditional β-AR agonists generally suppress T cells, they potentially enhance IL-17A production by Th17 cells. The effects of pharmacological drugs that count as biased agonists of AR like nebivolol are not completely understood. We investigated the impact of nebivolol on human memory CD4 T (Th1, Th2, Th17) cells and polarized naive Th17 cells, highlighting its potential for IL-17A suppression via a non-canonical β-AR cell signaling pathway.
The effects of nebivolol were tested on healthy human peripheral blood mononuclear cells, purified memory Th cells, and polarized naive Th17 cells activated with anti-CD3/anti-CD28/anti-CD2 ImmunoCult reagent. IFN-γ, IL-4, and IL-17A, which are primarily derived from Th1, Th2, and Th17 cells, respectively, were quantified by ELISA and flow cytometry. IL-10 was measured by ELISA. Gene expression of RORC, ADRB1, ADRB2, and ADRB3 was evaluated by qPCR. The ADRB2 gene was knocked out in memory Th cells using CRISPR/Cas9. Protein expression of phosphorylated serine133-CREB and phosphorylated NF-κB p65 was assessed by Western blot. Proliferation was assessed by fluorescent dye loading and flow cytometry.
Nebivolol treatment decreased IL-17A and IFN-γ secretion by activated memory Th cells and elevated IL-4 levels. Nebivolol reduced the proportion of IL-17A Th cells and downregulated RORC expression. Unlike the β-AR agonist terbutaline, nebivolol inhibited the shift of naive CD4 T cells toward the Th17 phenotype. IL-10 and the proliferation index remained unchanged. Nebivolol-treated β-knockout memory Th cells showed significant inhibition of β-AR-mediated signaling, evidenced by the absence of IL-17A suppression compared to controls. Phosphorylation of the NF-κB p65 subunit was inhibited by nebivolol, but CREB phosphorylation was not changed, suggesting a selective transcriptional control.
The findings demonstrate that nebivolol acts through a β-AR-mediated signaling pathway, as a distinctive anti-inflammatory agent capable of selectively shifting Th17 cells and suppressing the phosphorylation of NF-κB. This highlights nebivolol's potential for therapeutic interventions in chronic autoimmune conditions with elevated IL-17A levels.
肾上腺素能受体通过交感神经系统调节代谢、心血管和免疫功能。β-肾上腺素能受体（AR）作为高表达受体，对不同 T 细胞亚群的影响较为复杂，具体取决于配体的类型和所处的环境。传统的β-AR 激动剂通常会抑制 T 细胞，但它们可能会增强 Th17 细胞中 IL-17A 的产生。对于像奈必洛尔这样的 AR 部分激动剂类药物的药理作用还不完全清楚。我们研究了奈必洛尔对人类记忆性 CD4 T（Th1、Th2、Th17）细胞和极化的初始 Th17 细胞的影响，强调了其通过非典型β-AR 细胞信号通路抑制 IL-17A 的潜力。
用奈必洛尔处理健康人外周血单个核细胞、纯化的记忆性 Th 细胞和经抗 CD3/抗 CD28/抗 CD2 ImmunoCult 试剂激活的初始 Th17 细胞。通过 ELISA 和流式细胞术检测 IFN-γ、IL-4 和 IL-17A，它们分别主要来自 Th1、Th2 和 Th17 细胞。用 ELISA 法检测 IL-10。通过 qPCR 检测 RORC、ADRB1、ADRB2 和 ADRB3 的基因表达。用 CRISPR/Cas9 敲除记忆性 Th 细胞中的 ADRB2 基因。用 Western blot 检测磷酸化丝氨酸 133-CREB 和磷酸化 NF-κB p65 的蛋白表达。用荧光染料加载和流式细胞术检测增殖。
奈必洛尔治疗可降低激活的记忆性 Th 细胞中 IL-17A 和 IFN-γ 的分泌，并升高 IL-4 水平。奈必洛尔降低了 IL-17A Th 细胞的比例，并下调了 RORC 的表达。与β-AR 激动剂特布他林不同，奈必洛尔抑制了初始 CD4 T 细胞向 Th17 表型的转变。IL-10 和增殖指数保持不变。与对照组相比，奈必洛尔处理的β-AR 敲除记忆性 Th 细胞显示出明显的β-AR 介导的信号抑制，表明缺乏 IL-17A 的抑制作用。奈必洛尔抑制了 NF-κB p65 亚基的磷酸化，但 CREB 磷酸化未发生改变，表明存在选择性转录控制。
这些发现表明，奈必洛尔通过β-AR 介导的信号通路发挥作用，是一种独特的抗炎药物，能够选择性地改变 Th17 细胞并抑制 NF-κB 的磷酸化。这突出了奈必洛尔在治疗伴有升高的 IL-17A 水平的慢性自身免疫性疾病中的潜在作用。
