Retinal Pigment Epithelial Cells Suppress Phagolysosome Activation in Macrophages.
Retinal Pigment Epithelial Cells Suppress Phagolysosome Activation in Macrophages.
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DOI：
10.1167/iovs.16-21082
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发表时间：
2017-02-01
影响因子：
4.4
通讯作者：
Taylor AW
中科院分区：
文献类型：
--
作者：
Wang E;Choe Y;Ng TF;Taylor AW
眼睛是一个免疫特权微环境，已经适应了几种免疫调节机制来预防炎症。这些潜在机制之一是视网膜色素上皮细胞（RPE）改变巨噬细胞的吞噬作用。用健康小鼠和实验性自身免疫性葡萄膜炎（EAU）小鼠眼的RPE眼杯的条件培养液处理原代巨噬细胞吞噬pHrodo细菌生物颗粒。此外，从健康RPE眼杯的条件培养基中耗尽神经肽并用于处理吞噬巨噬细胞。测定来自健康和EAU RPE眼杯的条件培养基的IL-6，并将IL-6加入健康条件培养基中，并在EAU条件培养基中中和。用条件培养基处理巨噬细胞并测定荧光。对巨噬细胞进行成像，并测量相对于活性吞噬溶酶体的荧光强度。此外，使用荧光活力染料染色测定巨噬细胞。来自健康而不是EAU RPE眼杯的条件培养基抑制吞噬溶酶体活化。从健康RPE眼杯条件培养基中消耗神经肽α-黑素细胞刺激激素和神经肽Y导致巨噬细胞死亡。在EAU RPE眼杯中，条件培养基中的IL-6为0.96 ± 0.18 ng/mL，并且当中和时，条件培养基抑制吞噬溶酶体活化。健康的RPE通过可溶性分子，包括α-黑素细胞刺激激素和神经肽Y，抑制巨噬细胞中吞噬溶酶体的激活。在EAU中，由RPE产生的IL-6促进巨噬细胞中吞噬溶酶体的活化。这些结果表明，在健康条件下，RPE促进巨噬细胞中吞噬物质的改变途径，对抗原加工和清除有影响。
The eye is an immune-privileged microenvironment that has adapted several mechanisms of immune regulation to prevent inflammation. One of these potential mechanisms is retinal pigment epithelial cells (RPE) altering phagocytosis in macrophages. The conditioned media of RPE eyecups from eyes of healthy mice and mice with experimental autoimmune uveitis (EAU) were used to treat primary macrophage phagocytizing pHrodo bacterial bioparticles. In addition, the neuropeptides were depleted from the conditioned media of healthy RPE eyecups and used to treat phagocytizing macrophages. The conditioned media from healthy and EAU RPE eyecups were assayed for IL-6, and IL-6 was added to the healthy conditioned media, and neutralized in the EAU conditioned media. The macrophages were treated with the conditioned media and assayed for fluorescence. The macrophages were imaged, and the fluorescence intensity, relative to active phagolysosomes, was measured. Also, the macrophages were assayed using fluorescent viability dye staining. The conditioned media from healthy, but not from EAU RPE eyecups suppressed phagolysosome activation. Depletion of the neuropeptides alpha-melanocyte–stimulating hormone and neuropeptide Y from the healthy RPE eyecup conditioned media resulted in macrophage death. In the EAU RPE eyecup conditioned media was 0.96 ± 0.18 ng/mL of IL-6, and when neutralized the conditioned media suppressed phagolysosome activation. The healthy RPE through soluble molecules, including alpha-melanocyte–stimulating hormone and neuropeptide Y, suppresses the activation of the phagolysosome in macrophages. In EAU, the IL-6 produced by the RPE promotes the activation of phagolysosomes in macrophages. These results demonstrate that under healthy conditions, RPE promotes an altered pathway of phagocytized material in macrophages with implications on antigen processing and clearance.