Abstract

Innate immune cells, mainly monocytes/macrophages, upon their first encounter with a pathogen, form long-term nonspecific immunological memory, so-called “trained immunity.” In its formation, an important role is played by metabolites of the mevalonate pathway. The purpose of the study was to investigate the effect of mevalonate pathway modulators, mevalonate and zoledronate, on the formation of trained immunity in human and animal monocytes/macrophages. Human monocyte-like cells THP-1 and U-937 and peritoneal macrophages from BALB/c mice were used. Trained immunity was induced in vitro by incubating THP-1 and U-937 cells with inactivated mycobacteria of the bacillus Calmette–Guérin (BCG) vaccine strain for 24 and 72 h and in vivo by intraperitoneal injection of BCG to BALB/c mice and isolation of peritoneal macrophages on the seventh day after infection (lag phase). Cell hyperreactivity was assessed by the response to a second stimulus with bacterial lipopolysaccharide (LPS) and mevalanate or zoledranate in the presence or absence of LPS. The level of lactate, cytokines (IL-1β, TNF-α, IL-10), nitric oxide and glucose was evaluated in conditioned media from cells. It was found that monocyte-like THP-1 and U-937 cells respond differently as concerns the production of cytokines and lactate and the consumption of glucose to the BCG stimulus with or without the lag phase. Mevalonate and zoledronate alone or in combination with LPS also differentially stimulate cytokine secretion. The presence of the lag phase for human monocyte-like cells is essential for the level of cytokine production and glucose consumption. Peritoneal macrophages have been shown to increase the release of proinflammatory cytokines in response to LPS, mevalonate, and zoledronate. Mevalonate and zoledronate induce trained immunity in monocytes/macrophages.

中文翻译：

---

甲羟戊酸、唑来膦酸和 BCG 诱导对单核细胞/巨噬细胞表型的影响

摘要

先天免疫细胞，主要是单核细胞/巨噬细胞，在第一次遇到病原体时，会形成长期的非特异性免疫记忆，即所谓的“训练有素的免疫”。在其形成过程中，甲羟戊酸途径的代谢物发挥着重要作用。本研究的目的是研究甲羟戊酸途径调节剂甲羟戊酸和唑来膦酸对人类和动物单核细胞/巨噬细胞训练免疫形成的影响。使用人单核细胞样细胞 THP-1 和 U-937 以及来自 BALB/c 小鼠的腹膜巨噬细胞。通过将 THP-1 和 U-937 细胞与卡介苗 (BCG) 疫苗株的灭活分枝杆菌一起培养 24 和 72 小时，在体外诱导经过训练的免疫，并通过向 BALB/c 小鼠腹腔注射 BCG 并分离来诱导体内免疫感染后第七天的腹膜巨噬细胞（滞后期）。通过在存在或不存在 LPS 的情况下对细菌脂多糖 (LPS) 和甲羟戊酸或唑来膦酸的第二刺激的反应来评估细胞高反应性。在细胞条件培养基中评估乳酸、细胞因子（IL-1β、TNF-α、IL-10）、一氧化氮和葡萄糖的水平。研究发现，单核细胞样 THP-1 和 U-937 细胞在细胞因子和乳酸的产生以及葡萄糖的消耗方面对 BCG 刺激有或没有滞后期的反应不同。甲羟戊酸和唑来膦酸单独使用或与 LPS 联合使用也能不同程度地刺激细胞因子分泌。人单核细胞样细胞滞后期的存在对于细胞因子产生和葡萄糖消耗的水平至关重要。腹膜巨噬细胞已被证明可响应 LPS、甲羟戊酸和唑来膦酸而增加促炎细胞因子的释放。甲羟戊酸和唑来膦酸诱导单核细胞/巨噬细胞训练有素的免疫力。