MONOCYTE-INDUCED CYTOKINE EXPRESSION IN CULTURED HUMAN RETINAL-PIGMENT EPITHELIAL-CELLS
MONOCYTE-INDUCED CYTOKINE EXPRESSION IN CULTURED HUMAN RETINAL-PIGMENT EPITHELIAL-CELLS
复制标题
DOI：
10.1016/s0014-4835(05)80068-5
复制
发表时间：
1995-05-01
影响因子：
3.4
通讯作者：
CIANCIOLO, GJ
中科院分区：
文献类型：
--
作者：
JAFFE, GJ;ROBERTS, WL;CIANCIOLO, GJ
单核细胞和视网膜色素上皮细胞在增殖性玻璃体视网膜病变和某些类型的葡萄膜炎的眼膜中密切相关。本研究的目的是确定单核细胞是否调节视网膜色素上皮细胞中细胞因子的表达，如果是这样，以确定单核细胞的产品负责这种效果。将培养的人视网膜色素上皮细胞暴露于来自未刺激的人单核细胞或来自用脂多糖预刺激的单核细胞的不同浓度的单核细胞条件培养基1-48小时。通过逆转录聚合酶链反应测定白细胞介素-1 β、白细胞介素-6、白细胞介素-8、黑色素瘤生长刺激活性/gro α和γ、巨噬细胞集落刺激因子、转化生长因子-β 2、碱性成纤维细胞生长因子和激活素β A链的mRNA表达。通过ELISA在RPE条件培养基中测量所选细胞因子、白细胞介素-1 β、白细胞介素-6、白细胞介素-8、巨噬细胞集落刺激因子和转化生长因子-β 2的蛋白分泌。视网膜色素上皮细胞组成性表达白细胞介素-6、巨噬细胞集落刺激因子、转化生长因子-β 2、碱性成纤维细胞生长因子和激活素β A链的mRNA。白细胞介素-1 β、黑色素瘤生长刺激活性/gro α和γ以及白细胞介素-8在基础条件下不表达。除碱性成纤维细胞生长因子（bFGF）和转化生长因子β 2（TGF-β 2）外，刺激的单核细胞条件培养液对所有细胞因子的mRNA表达均有明显的诱导作用，且呈剂量和时间依赖性。刺激的单核细胞条件培养基还诱导白细胞介素-6、白细胞介素-8、巨噬细胞集落刺激因子和转化生长因子-β 2的时间依赖性增加，但不诱导白细胞介素-1 β蛋白分泌（所有时间点p < 0.05）。中和抗体白细胞介素-1 β，或肿瘤坏死因子α，但不是白细胞介素-1 α，显着降低细胞因子mRNA的表达诱导刺激单核细胞条件培养基。所有三种中和抗体的组合几乎完全消除了所研究的所有细胞因子的单核细胞诱导的mRNA表达和蛋白质产生。活化的单核细胞分泌的产物的异质混合物一起强烈诱导多种细胞因子在人视网膜色素上皮细胞的表达。大多数（如果不是全部的话）诱导作用可以由白细胞介素-lp和肿瘤坏死因子a解释。由于细胞因子与增殖性玻璃体视网膜病变和葡萄膜炎有关，因此RPE细胞的单核细胞介导的细胞因子表达可能导致这些疾病的发生和延续。
Monocytes and retinal pigment epithelial cells are intimately associated in membranes of eyes with proliferative vitreoretinopathy and in certain types of uveitis. The goal of this study was to determine whether monocytes modulate cytokine expression in retinal pigment epithelial cells, and if so, to identify the monocyte products responsible for this effect. Cultured human retinal pigment epithelial cells were exposed to Varying concentrations of monocyte-conditioned medium from unstimulated human monocytes for 1-48 hr, or from monocytes prestimulated with lipopolysaccharide. mRNA expression of interleukin-1 beta, interleukin-6, interleukin-8, melanoma growth stimulating activity/gro alpha and gamma, macrophage colony stimulating factor, transforming growth factor-beta 2, basic fibroblast growth factor and activin beta A chain was determined by reverse transcription polymerase chain reaction. Protein secretion of selected cytokines, interleukin-1 beta, interleukin-6, interleukin-8, macrophage colony stimulating factor and transforming growth factor-beta 2 was measured in RPE-conditioned medium by ELISA. Retinal pigment epithelial cells constitutively expressed mRNA for interleukin-6, macrophage colony stimulating factor, transforming growth factor-beta 2, basic fibroblast growth factor and activin beta A chain. Interleukin-1 beta, melanoma growth stimulating activity/gro alpha and gamma and interleukin-8 were not expressed under basal conditions. Stimulated monocyte-conditioned medium markedly induced mRNA of all cytokines except basic fibroblast growth factor and transforming growth factor-beta 2 in a dose- and time-dependent manner, Unstimulated monocyte-conditioned medium was a less potent inducing agent, but still enhanced mRNA expression of interleukin-6, interleukin-8 and melanoma growth stimulating activity/gro a. Stimulated monocyte-conditioned medium also induced a time-dependent increase in interleukin-6, Interleukin-8, macrophage colony stimulation factor and transforming growth factor-beta 2, but not interleukin-1 beta protein secretion (p < 0.05 for all time points). Neutralizing antibodies to interleukin-1 beta, or tumour necrosis factor alpha, but not interleukin-1 alpha, significantly reduced cytokine mRNA expression induced by stimulated monocyte-conditioned medium. the combination of all three neutralizing antibodies almost entirely eliminated monocyte-induced mRNA expression and protein production of all cytokines studied. Activated monocytes secrete a heterogeneous mixture of products that together strongly induce expression of multiple cytokines in human retinal pigment epithelial cells. Most if not all of the inducing effect can be accounted for by interleukin-lp and tumour necrosis factor a. Because cytokines have been implicated in proliferative vitreoretinopathy and uveitis, monocyte-mediated cytokine expression by RPE cells may serve to initiate and perpetuate these diseases.