METHODS: CD4+ T cells that were isolated from blood samples from healthy individuals and from patients with type 1 diabetes (T1D) were activated in vitro. We carried out metabolic assays, multiple proximity extension assay (PEA), ELISA, qPCR, immunoblotting, immunofluorescence staining, flow cytometry analysis, MS-based proteomics, as well as electrophysiology and live-cell Ca2+ imaging.
RESULTS: We demonstrate that GABA-mediated reduction of metabolic activity and the release of inflammatory proteins, including IFNγ and IL-10, were abolished in human CD4+ T cells from healthy individuals and patients with T1D when the glucose concentration was elevated above levels typically observed in healthy people. Insulin increased GABAA receptor-subunit ρ2 expression, enhanced the GABAA receptors-mediated currents and Ca2+ influx. GABA decreased, whereas insulin sustained, hexokinase activity and glycolysis in a glucose concentration-dependent manner.
CONCLUSIONS: These findings support that metabolic factors, such as glucose and insulin, influence the GABA-mediated immunomodulation of human primary T cells effector functions.
BACKGROUND: The Swedish Children\'s Diabetes Foundation, The Swedish Diabetes Foundation, The Swedish Research Council 2018-02952, EXODIAB, The Ernfors Foundation, The Thurings Foundation and the Science for Life Laboratory.
方法:从健康个体和1型糖尿病(T1D)患者的血液样本中分离的CD4+T细胞在体外被激活。我们进行了代谢测定,多重邻近延伸测定(PEA),ELISA,qPCR,免疫印迹,免疫荧光染色,流式细胞术分析,基于MS的蛋白质组学,以及电生理学和活细胞Ca2+成像。
结果:我们证明了GABA介导的代谢活性降低和炎症蛋白的释放,当葡萄糖浓度升高到高于健康人中通常观察到的水平时,包括IFNγ和IL-10在来自健康个体和患有T1D的患者的人CD4+T细胞中被废除。胰岛素增加GABAA受体亚基ρ2表达,增强了GABAA受体介导的电流和Ca2流入。GABA下降,而胰岛素持续,己糖激酶活性和糖酵解呈葡萄糖浓度依赖性。
结论:这些发现支持代谢因素,如葡萄糖和胰岛素,影响GABA介导的人原代T细胞效应功能的免疫调节。
背景:瑞典儿童糖尿病基金会,瑞典糖尿病基金会TheSwedishResearchCouncil2018-02952,EXODIAB,Ernfors基金会,图林斯基金会和生命科学实验室。