Immunoglobulin Activation of Fibroblasts

成纤维细胞的免疫球蛋白激活

• 批准号: 6824455
• 负责人: TERRY J SMITH
• 金额: $ 32.12万
• 依托单位: LUNDQUIST INSTITUTE FOR BIOMEDICAL INNOVATION AT HARBOR-UCLA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6824455
• 关键词: Graves disease; SCID mouse; antibody; biological signal transduction; chemokine; clinical research; enzyme linked immunosorbent assay; fibroblasts; gene induction /repression; genetic regulation; genetic transcription; genetic translation; growth factor receptors; human subject; human tissue; immunocytochemistry; immunoglobulin G; in situ hybridization; insulinlike growth factor; ligands; longitudinal human study; molecular pathology; northern blottings; phosphorylation; receptor expression; western blottings; xenotransplantation

DESCRIPTION (provided by applicant): Fibroblast activation plays an important role in initiating the inflammatory response. Phenotypic attributes of fibroblasts from specific anatomic regions are thought to underlie the peculiar pattern of manifestations associated with certain diseases. Other fibroblast characteristics appear global, such as the expression of chemoattractants including IL-16, a CD4-specific ligand and RANTES, a C-C chemokine. We have found that fibroblasts from patients with the Graves' disease (GD), when treated with their IgGs (GD-IgG), become activated and express high levels of IL-16 and RANTES and in so doing provoke the migration of T cells. Control fibroblasts from donors without autoimmune disease fail to respond to these IgGs. GD-IgGs, which are rare in control sera, appear to be binding to the insulin-like growth factor-1 receptor (IGF-1R) displayed on fibroblasts. We hypothesize that IGF-1R represents an important activational self-antigen. Its ligation with GD-IgG up-regulates chemoattractant expression in fibroblasts. Further, we hypothesize that the IGF-1R display by disease-derived cultures in some way differs from that on control fibroblasts. These inductions could underlie T cell infiltration in GD. We now propose the following studies to test our central hypothesis. Specific aims: (1) To define the mechanisms involved in the up-regulation by anti-IGF-1R Abs (i.e. GD-IgG) of IL-16 and RANTES expression in GD- fibroblasts by assessing transcriptional and translational regulation and cell signaling through IGF-1R. (2) To determine whether fibroblasts from patients with GD express functionally different IGF-1R from control fibroblasts, we will perform IGF-1 binding studies, northern and western blot analysis of IGF-1R alpha and beta subunit levels and examine IGF-1R tyrosine phosphorylation. (3) To determine whether serum and tissue levels of IL-16 and RANTES are higher in GD by performing ELISA, in situ hybridization and immunohistochemical studies. (4) To determine whether GD-IgG induces chemoattractant expression in vivo in xenotransplanted human orbital tissue from patients with GD in SCID mice. We believe that important insights into disease pathogenesis and identification of novel therapeutic targets will emerge from these studies.

描述（由申请人提供）：成纤维细胞活化在引发炎症反应中发挥重要作用。来自特定解剖区域的成纤维细胞的表型属性被认为是与某些疾病相关的特殊表现模式的基础。其他成纤维细胞特征似乎是全局性的，例如趋化剂的表达，包括 IL-16（一种 CD4 特异性配体）和 RANTES（一种 C-C 趋化因子）。我们发现，格雷夫斯病 (GD) 患者的成纤维细胞在接受 IgG (GD-IgG) 治疗后会被激活并表达高水平的 IL-16 和 RANTES，从而引发 T 细胞迁移。来自没有自身免疫性疾病的供体的对照成纤维细胞无法对这些 IgG 产生反应。 GD-IgG 在对照血清中很少见，似乎与成纤维细胞上的胰岛素样生长因子 1 受体 (IGF-1R) 结合。我们假设 IGF-1R 代表一种重要的激活性自身抗原。它与 GD-IgG 的连接可上调成纤维细胞中趋化剂的表达。此外，我们假设疾病来源的培养物显示的 IGF-1R 在某种程度上不同于对照成纤维细胞。这些诱导可能是 GD 中 T 细胞浸润的基础。我们现在提出以下研究来检验我们的中心假设。具体目标：(1) 通过评估转录和翻译调节以及细胞信号传导，确定抗 IGF-1R Abs（即 GD-IgG）上调 GD-成纤维细胞中 IL-16 和 RANTES 表达的机制。 IGF-1R。 (2) 为了确定 GD 患者的成纤维细胞是否表达与对照成纤维细胞功能上不同的 IGF-1R，我们将进行 IGF-1 结合研究、IGF-1R α 和 β 亚基水平的 Northern 和 Western 印迹分析，并检查 IGF-1R 酪氨酸磷酸化。 (3)通过ELISA、原位杂交和免疫组化研究确定GD患者血清和组织中IL-16和RANTES的水平是否较高。 (4) 确定GD-IgG是否在SCID小鼠中的GD患者异种移植人眼眶组织中诱导体内趋化剂表达。我们相信这些研究将产生对疾病发病机制和新治疗靶点识别的重要见解。