Department of Urology, University of Florida, Gainesville, FL, United States.
Department of Pathology, Immunology and Laboratory Medicine, College of Medicine, University of Florida, Gainesville, FL, United States.
Front Immunol. 2018 Aug 22;9:1863. doi: 10.3389/fimmu.2018.01863. eCollection 2018.
A number of hyperoxaluric states have been associated with calcium oxalate (CaOx) deposits in the kidneys. In animal models of stone disease, these crystals interact with circulating monocytes that have migrated into the kidney as part of innate immunity. Similarly, macrophages surround CaOx crystals in kidneys of patients excreting high levels of oxalate. We investigate the effect of this exposure and subsequent human immunological response .
Primary human monocytes were collected from healthy donors and exposed to CaOx, potassium oxalate, and zinc oxalate (ZnOx). Cytokine production was measured with a multiplex ELISA. Quantitative reverse transcription-polymerase chain reaction was done to validate the mRNA profile expression. M1 macrophage phenotype was confirmed with immunofluorescence microscopy.
Both primary monocytes and THP-1 cells, a human monocytic cell line, respond strongly to CaOx crystals in a dose-dependent manner producing TNF-α, IL-1β, IL-8, and IL-10 transcripts. Exposure to CaOx followed by 1 h with LPS had an additive effect for cytokine production compared to LPS alone, however, LPS followed by CaOx led to significant decrease in cytokine production. Supernatants taken from monocytes were previously exposed to CaOx crystals enhance M2 macrophage crystal phagocytosis. CaOx, but not potassium or ZnOx, promotes monocyte differentiation into inflammatory M1-like macrophages.
In our experiment, human monocytes were activated by CaOx and produced inflammatory cytokines. Monocytes recognized CaOx crystals through a specific mechanism that can enhance or decrease the innate immune response to LPS. CaOx promoted M1 macrophage development. These results suggest that monocytes have an important role promoting CaOx-induced inflammation.
许多高草酸尿症与肾脏中的草酸钙 (CaOx) 沉积有关。在结石病的动物模型中，这些晶体与循环单核细胞相互作用，单核细胞作为先天免疫的一部分迁移到肾脏。同样，在排泄高草酸的患者的肾脏中，巨噬细胞围绕着 CaOx 晶体。我们研究了这种暴露及其随后的人体免疫反应的影响。
从健康供体中收集原代人单核细胞，并将其暴露于 CaOx、草酸钾和草酸锌 (ZnOx) 下。使用多重 ELISA 测量细胞因子产生。通过定量逆转录聚合酶链反应验证 mRNA 谱表达。通过免疫荧光显微镜确认 M1 巨噬细胞表型。
原代单核细胞和人单核细胞系 THP-1 细胞均对 CaOx 晶体表现出强烈的剂量依赖性反应，产生 TNF-α、IL-1β、IL-8 和 IL-10 转录本。与单独 LPS 相比，暴露于 CaOx 后再用 LPS 1 小时会产生细胞因子产生的附加效应，然而，LPS 后再用 CaOx 会导致细胞因子产生显著减少。先前暴露于 CaOx 晶体的单核细胞上清液增强了 M2 巨噬细胞对晶体的吞噬作用。CaOx 但不是草酸钾或 ZnOx 促进单核细胞分化为炎症性 M1 样巨噬细胞。
在我们的实验中，人单核细胞被 CaOx 激活并产生炎症细胞因子。单核细胞通过一种特定的机制识别 CaOx 晶体，这种机制可以增强或减弱对 LPS 的先天免疫反应。CaOx 促进了 M1 巨噬细胞的发展。这些结果表明单核细胞在促进 CaOx 诱导的炎症中起着重要作用。
