Calcium Oxalate Stone Fragment and Crystal Phagocytosis by Human Macrophages.
Calcium Oxalate Stone Fragment and Crystal Phagocytosis by Human Macrophages.
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DOI：
10.1016/j.juro.2015.11.048
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发表时间：
2016-04
期刊：
影响因子：
--
通讯作者：
Khan SR
中科院分区：
文献类型：
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作者：
Kusmartsev S;Dominguez-Gutierrez PR;Canales BK;Bird VG;Vieweg J;Khan SR
在小鼠和人类高尿酸条件下，可以看到巨噬细胞周围的肾草酸钙（CaOx）晶体沉积。我们假设巨噬细胞在降解和破坏这些沉积物中发挥作用，并研究了巨噬细胞暴露于人肾结石和无机晶体时的炎症反应和吞噬机制。通过用重组人M-CSF或GM-CSF处理6天，将人单核细胞分化成静息的、完全分化的巨噬细胞。在通过流式细胞术确认表型后，将巨噬细胞暴露于无菌人CaOx结石或CaOx晶体的片段20小时。测定晶体摄取，并使用抗体阵列分析上清液细胞因子和趋化因子谱。qRT-PCR用于验证mRNA谱表达。在直视荧光显微镜下，发现激活的人巨噬细胞包围结石碎片和合成晶体，并在涉及网格蛋白介导的内吞作用和吞噬作用的逐步过程中破坏它们。巨噬细胞释放炎性级联反应，包括趋化因子CCL 2、CCL 3、白细胞介素-1受体拮抗剂（IL-1 ra）、补体成分C5/C5 a和IL-8。对结石和晶体材料的反应模式取决于巨噬细胞表型和活化状态。在我们的体外研究中，M-CSF分化的巨噬细胞比GM-CSF处理的巨噬细胞显示出更大的吞噬晶体沉积物的能力。在网格蛋白介导的内吞作用后，巨噬细胞释放了许多对炎症免疫反应至关重要的细胞因子，这表明组织巨噬细胞通过清除和消化肾间质晶体沉积物在预防肾结石疾病中发挥重要作用。
In murine and human hyperoxaluric conditions, macrophages can be seen surrounding renal calcium oxalate (CaOx) crystal deposits. We hypothesize that macrophages play a role in degrading and destroying these deposits and investigated inflammatory response and phagocytic mechanisms when macrophages are exposed to human kidney stones and inorganic crystals. Human monocytes were differentiated into resting, fully-differentiated macrophages by treating with recombinant human M-CSF or GM-CSF for 6 days. After confirming phenotype by flow cytometry, macrophages were exposed for 20 hours to fragments of sterile human CaOx stones or CaOx crystals. Crystal uptake was determined, and supernatant cytokine and chemokine profiles were analyzed using antibody arrays. qRT-PCR was used to validate mRNA profile expression. Under direct-vision fluorescent microscopy, activated human macrophages were noted to surround both stone fragments and synthesized crystals and destroy them in a step-by-step process that involved clathrin-mediated endocytosis and phagocytosis. An inflammatory cascade was released by macrophages, including chemokines CCL2, CCL3, interleukin-1 receptor antagonist (IL-1ra), complement component C5/C5a and IL-8. The response patterns to stone and crystal material was dependent on macrophage phenotype and activation status. In our in vitro study, macrophages differentiated with M-CSF displayed a greater ability to phagocytize crystal deposits than those treated with GM-CSF. Following clathrin-mediated endocytosis, macrophages released a number of cytokines crucial for inflammatory immune response, suggesting that tissue macrophages play an important role in preventing kidney stone disease by removing and digesting interstitial renal crystal deposits.