Education-dependent activation of glycolysis promotes the cytolytic potency of licensed human natural killer cells
Education-dependent activation of glycolysis promotes the cytolytic potency of licensed human natural killer cells
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DOI：
10.1016/j.jaci.2018.06.047
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发表时间：
2019-01-01
影响因子：
14.2
通讯作者：
Lee, Dean A.
中科院分区：
文献类型：
--
作者：
Schafer, Jolie R.;Salzillo, Travis C.;Lee, Dean A.
背景资料：自然杀伤（NK）细胞教育导致获得许可的NK细胞对缺失的自身靶点具有增强的效应器功能的机制尚不清楚。目的：我们试图确定获得许可的人类NK细胞增强功能的潜在机制。方法：我们使用来自杀伤免疫球蛋白样受体（KIR）/HLA基因分型供体的扩增的人NK细胞，其分选为单KIR +细胞以产生经许可和未经许可的NK细胞的纯群体。我们在受体交联前后对这些细胞进行了蛋白质组学和基因表达分析，并在干扰选定的代谢途径前后进行了功能和代谢分析。我们使用新鲜分离和分选的NK细胞从外周blood.Results：我们证实，许可的人NK细胞在外周血中的数量更大，在体外增殖比未经许可的NK细胞。使用高通量蛋白质分析，我们发现未经刺激的许可NK细胞具有糖酵解酶丙酮酸激酶肌肉同工酶M2的增加的表达，并且在KIR交联后具有代谢调节剂p38-α和5 '腺苷一磷酸活化的蛋白激酶α的增加的磷酸化。在细胞因子扩增和激活后，未经许可的NK细胞仅依赖于线粒体呼吸来实现细胞溶解功能，而经许可的NK细胞表现出朝向糖酵解和唾液酸依赖性β-氨基醇解的代谢重编程，导致糖酵解代谢物的积累和谷氨酸的耗尽。因此，需要阻断糖酵解和呼吸依赖性呼吸来抑制持牌NK细胞的细胞毒性。结论：总的来说，我们的数据支持武装模式的教育，其中增强糖酵解持牌NK细胞支持增殖和细胞毒性能力。
Background: The mechanism by which natural killer (NK) cell education results in licensed NK cells with heightened effector function against missing self-targets is not known.Objectives: We sought to identify potential mechanisms of enhanced function in licensed human NK cells.Methods: We used expanded human NK cells from killer immunoglobulin-like receptor (KIR)/HLA-genotyped donors sorted for single-KIR+ cells to generate pure populations of licensed and unlicensed NK cells. We performed proteomic and gene expression analysis of these cells before and after receptor cross-linking and performed functional and metabolic analysis before and after interference with selected metabolic pathways. We verified key findings using freshly isolated and sorted NK cells from peripheral blood.Results: We confirmed that licensed human NK cells are greater in number in peripheral blood and proliferate more in vitro than unlicensed NK cells. Using high-throughput protein analysis, we found that unstimulated licensed NK cells have increased expression of the glycolytic enzyme pyruvate kinase muscle isozyme M2 and after KIR cross-linking have increased phosphorylation of the metabolic modulators p38-alpha and 5 ' adenosine monophosphate-activated protein kinase a. After cytokine expansion and activation, unlicensed NK cells depended solely on mitochondrial respiration for cytolytic function, whereas licensed NK cells demonstrated metabolic reprogramming toward glycolysis and mitochondrial-dependent glutaminolysis, leading to accumulation of glycolytic metabolites and depletion of glutamate. As such, blocking both glycolysis and mitochondrial-dependent respiration was required to suppress the cytotoxicity of licensed NK cells.Conclusions: Collectively, our data support an arming model of education in which enhanced glycolysis in licensed NK cells supports proliferative and cytotoxic capacity.