Takizawa T, Nishinarita S, Kitamura N, Hayakawa J, Kang H, Tomita Y, Mitamura K, Yamagami K, Horie T
First Department of Internal Medicine, Nihon University School of Medicine, Tokyo, Japan.
Clin Exp Immunol. 1995 Aug;101(2):376-82. doi: 10.1111/j.1365-2249.1995.tb08367.x.
The effect of fibronectin on IL-1 alpha, IL-1 beta, tumour necrosis factor-alpha (TNF-alpha), and IL-6 production was investigated with cultured monocytes isolated from human peripheral blood. Monokine concentrations were determined by both ELISA and bioassay. Fibronectin markedly stimulated the secretion of IL-1 alpha, IL-1 beta, TNF-alpha and IL-6 from cultured monocytes in a dose-dependent manner, with the maximal effect apparent within 24 h. Northern blot analysis revealed a marked increase in the abundance of mRNA specific for each monokine on exposure of monocytes to fibronectin. Monoclonal antibodies to the alpha chain of very late antigen (VLA)-5, the beta 1 integrin, the alpha chain of Mac-1, and the beta 2 integrin, as well as the synthetic peptide of GRGDSP (which corresponds to the cell-binding domain of fibronectin), inhibited (> 50%) fibronectin-induced monokine production. Monoclonal antibodies to the alpha chain of VLA-4, and the alpha chain of LFA-1, as well as the synthetic peptide CS-1 (which corresponds to the alternatively spliced connecting segment of fibronectin) and the control peptide GRADSP, had no inhibitory effect on monokine production. A MoAb, R60, that recognizes an epitope of the fibronectin molecule that includes the RGD sequence, inhibited monokine production, whereas the MoAb Y16, which recognizes another epitope of fibronectin not including RGD, did not. These results indicate that fibronectin-induced production of IL-1 alpha, IL-1 beta, TNF-alpha and IL-6 from cultured monocytes is mediated predominantly by interaction of the cell-binding domain of fibronectin with VLA-5, although Mac-1 also may contribute to this effect of fibronectin. Our results indicate that the interaction of fibronectin with integrins may contribute to the cytokine network in inflammatory response.
利用从人外周血中分离的培养单核细胞，研究了纤连蛋白对白细胞介素-1α（IL-1α）、白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）和白细胞介素-6产生的影响。通过酶联免疫吸附测定法（ELISA）和生物测定法测定单核因子浓度。纤连蛋白以剂量依赖方式显著刺激培养单核细胞分泌IL-1α、IL-1β、TNF-α和IL-6，最大效应在24小时内明显。Northern印迹分析显示，单核细胞暴露于纤连蛋白后，每种单核因子特异性mRNA的丰度显著增加。针对极晚期抗原（VLA）-5的α链、β1整合素链、Mac-1的α链和β2整合素链的单克隆抗体，以及GRGDSP合成肽（对应于纤连蛋白的细胞结合结构域），抑制（>50%）纤连蛋白诱导的单核因子产生。针对VLA-4的α链和淋巴细胞功能相关抗原-1（LFA-1）的α链的单克隆抗体，以及合成肽CS-1（对应于纤连蛋白的可变剪接连接段）和对照肽GRADSP，对单核因子产生无抑制作用。识别包含RGD序列的纤连蛋白分子表位的单克隆抗体R60抑制单核因子产生，而识别不包含RGD的纤连蛋白另一表位的单克隆抗体Y16则无此作用。这些结果表明，纤连蛋白诱导培养单核细胞产生IL-1α、IL-1β、TNF-α和IL-6主要是由纤连蛋白的细胞结合结构域与VLA-5相互作用介导的，尽管Mac-1也可能有助于纤连蛋白的这一作用。我们的结果表明，纤连蛋白与整合素的相互作用可能在炎症反应的细胞因子网络中起作用。
