Retinal microglia differentially express phenotypic markers of antigen-presenting cells in vitro.
Retinal microglia differentially express phenotypic markers of antigen-presenting cells in vitro.
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DOI：
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发表时间：
1999-12
影响因子：
4.4
通讯作者：
Takashi Matsubara;Takashi Matsubara;G. Pararajasegaram;Guey-Shuang Wu;Narsing A. Rao
中科院分区：
文献类型：
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作者：
Takashi Matsubara;Takashi Matsubara;G. Pararajasegaram;Guey-Shuang Wu;Narsing A. Rao
目的分离培养新生Lewis大鼠视网膜小胶质细胞，研究巨噬细胞集落刺激因子（M-CSF）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）和干扰素-γ（IFN-γ）对小胶质细胞抗原呈递所需辅助分子表达的影响。方法从新生Lewis大鼠中分离视网膜小胶质细胞，并在补充有M-CSF或GM-CSF的培养基中培养。使用抗巨噬细胞补体受体 3 (OX42) 或抗单核巨噬细胞 (ED1) 和 DiI-ac-低密度脂蛋白 (LDL) 摄取的免疫组织化学测试来识别小胶质细胞。使用以下抗体之一，通过荧光激活细胞分选仪分析在不同条件（M-CSF、GM-CSF 和 M-CSF 加 IFN-γ）下培养的小胶质细胞对辅助分子表达的影响：抗 OX3、抗 OX6、抗大鼠细胞间粘附分子 (ICAM)-1、抗大鼠 B7-1 或抗大鼠 B7-2。结果培养的视网膜小胶质细胞巨噬细胞相关抗原（ED1和OX42）呈阳性，并且还显示出对LDL的摄取。此外，ICAM-1和B7-2在这些细胞上组成型表达，并且MHC II类和B7-1在IFN-γ刺激后也表达。结论 在体外，视网膜小胶质细胞表达有效抗原呈递给 CD4 阳性 T 细胞所需的分子。这些发现表明，小胶质细胞可能在局部抗原呈递中发挥作用，特别是当它们暴露于 IFN-γ 时。
PURPOSE Retinal microglial cells of newborn Lewis rats were isolated and cultured, and the effect of macrophage colony-stimulating factor (M-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and interferon-gamma (IFN-gamma) on microglial expression of the accessory molecules required for antigen presentation were studied. METHODS Retinal microglia were isolated from newborn Lewis rats and cultured in media supplemented with either M-CSF or GM-CSF. Immunohistochemical tests using anti-macrophage complement receptor 3 (OX42) or anti-monocyte-macrophage (ED1) and DiI-ac-low-density lipoprotein (LDL) uptake were used to identify microglia. The effect on accessory molecule expression of microglial cells cultured under varying conditions (M-CSF, GM-CSF, and M-CSF plus IFN-gamma) was analyzed by fluorescence-activated cell sorter, using one of the following antibodies: anti-OX3, anti-OX6, anti-rat intercellular adhesion molecule (ICAM)-1, anti-rat B7-1, or anti-rat B7-2. RESULTS The cultured retinal microglia were positive for macrophage-related antigens (ED1 and OX42) and also showed uptake of LDL. Furthermore, ICAM-1 and B7-2 were expressed constitutively on these cells, and MHC class II and B7-1 were also expressed after IFN-gamma stimulation. CONCLUSIONS In vitro, the retinal microglia express the molecules required for effective antigen presentation to CD4-positive T cells. These findings suggest that microglia may play a role in local antigen presentation, especially when they are exposed to IFN-gamma.