Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
Nagasaki University Graduate School of Biomedical Sciences and Nagasaki University Hospital, Nagasaki, Japan.
Arthritis Rheumatol. 2019 May;71(5):766-772. doi: 10.1002/art.40785. Epub 2019 Mar 20.
To clarify the significance of immunometabolism in systemic lupus erythematosus (SLE), and to determine the effect of calcium/calmodulin-dependent protein kinase 4 (CaMK4) on T cell metabolism.
Metabolomic profiling was performed using capillary electrophoresis mass spectrometry in naive T cells from MRL/lpr mice treated with anti-CD3/CD28 antibodies in the absence or presence of a CaMK4 inhibitor (KN-93). The expression of GLUT1 and CaMK4 in CD4+ T cells from healthy controls (n = 16), patients with inactive SLE (n = 13), and patients with active SLE (n = 14) was examined by flow cytometry and quantitative polymerase chain reaction. In vitro experiments were performed to determine the effect of KN-93 on the expression of GLUT1 during Th17 cell differentiation in T cells from patients with SLE.
CaMK4 inhibition significantly decreased the levels of glycolytic intermediates such as glucose-6-phosphate, fructose-6-phosphate, fructose-1,6-diphosphate, pyruvate, and lactate (P < 0.05), whereas it did not affect the levels of the pentose phosphate pathway intermediates such as 6-phospho-d-gluconate, ribulose-5-phosphate, ribose-5-phosphate, and phosphoribosyl pyrophosphate. The expression levels of GLUT1 and CaMK4 in effector memory CD4+ T cells were significantly higher in patients with active SLE compared to healthy controls (P < 0.01 and P < 0.05, respectively) and patients with inactive SLE (P < 0.05 and P < 0.01, respectively). A functional analysis revealed that CaMK4 inhibition decreased the expression of GLUT1 during Th17 cell differentiation (P < 0.01), followed by a reduction of interleukin-17 (IL-17) production (P < 0.05).
The results of the study indicate that the activity of CaMK4 could be responsible for glycolysis, which contributes to the production of IL-17, and CaMK4 may contribute to aberrant expression of GLUT1 in T cells from patients with active SLE.
阐明免疫代谢在系统性红斑狼疮（SLE）中的意义，并确定钙/钙调蛋白依赖性蛋白激酶 4（CaMK4）对 T 细胞代谢的影响。
使用毛细管电泳-质谱联用技术对未经处理以及经 CaMK4 抑制剂 KN-93 处理的 MRL/lpr 小鼠的初始 T 细胞进行代谢组学分析。通过流式细胞术和实时聚合酶链反应检测来自健康对照者（n=16）、非活动期 SLE 患者（n=13）和活动期 SLE 患者（n=14）的 CD4+T 细胞中 GLUT1 和 CaMK4 的表达。在体外实验中，研究了 KN-93 对 SLE 患者 T 细胞中 Th17 细胞分化过程中 GLUT1 表达的影响。
CaMK4 抑制显著降低了糖酵解中间产物的水平，如葡萄糖-6-磷酸、果糖-6-磷酸、果糖-1,6-二磷酸、丙酮酸和乳酸（P<0.05），而对戊糖磷酸途径中间产物的水平没有影响，如 6-磷酸-d-葡萄糖酸、核酮糖-5-磷酸、核糖-5-磷酸和磷酸核糖焦磷酸。与健康对照者（P<0.01 和 P<0.05）和非活动期 SLE 患者（P<0.05 和 P<0.01）相比，活动期 SLE 患者效应记忆 CD4+T 细胞中 GLUT1 和 CaMK4 的表达水平明显更高（P<0.01 和 P<0.05）。功能分析显示，CaMK4 抑制降低了 Th17 细胞分化过程中 GLUT1 的表达（P<0.01），随后减少了白细胞介素-17（IL-17）的产生（P<0.05）。
研究结果表明，CaMK4 的活性可能与糖酵解有关，而糖酵解有助于 IL-17 的产生，CaMK4 可能导致活动期 SLE 患者 T 细胞中 GLUT1 的异常表达。
