Cytokines And Growth Factors In Autoimmune Diseases

自身免疫性疾病中的细胞因子和生长因子

• 批准号: 6814510
• 负责人: Ashok B. KULKARNI
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6814510
• 关键词: arthritis; autoimmune disorder; cytokine; histocompatibility antigens; immunopathology therapy; immunotherapy; inflammation; laboratory mouse; nonhuman therapy evaluation; systemic lupus erythematosus; transforming growth factors

Cytokines and growth factors play critical roles in normal homeostasis of immune functions. TGF-?1 is the dominant regulator of inflammatory responses. We have earlier generated TGF-?1-/- mice that exhibit multifocal inflammation associated with increased adhesion of leukocytes to endothelium, aberrant expression of MHC class-I and -II antigens, and autoimmune manifestations similar to Sjogren syndrome. Moreover, depletion of CD8 cells ameliorated health status of TGF-?1 null mice. To delineate role of TGF-?1 in aberrant MHC expression, TGF-?1 null mice were generated in the MHC-I and MHC-II deficient backgrounds. TGF-?1 X MHC-I null mice exhibit increased longevity associated with reduced inflammation, diminished autoimmune manifestations and significantly elevated myelopoiesis. Although the TGF-?1 X MHC-II mice exhibited lack of inflammation and autoimmune response, they showed neonatal mortality associated with increased myelopoiesis and metaplasia. These results indicate a dominant role of TGF-?1 in leukocyte maturation and function. To assess the therapeutic potential of circulating levels of active TGF-?1, we generated mice with endocrine expression of active TGF-?1 on a TGF-?1 null background (TGF-?1(-/-/TG)) by crossing TGF-?1(+/-) mice with transgenic mice (TG) that express recombinant TGF-?1 specifically in the liver and secrete it in the blood. The TGF-_1(-/-/TG) mice exhibit a survival profile similar to the TGF-?1 (-/-) mice indicating a failure to rescue the lethal phenotype. However, serum TGF-?1 levels in theTGF-?1 (-/-/TG) mice were restored to normal levels with expression in all the tissues notably in the kidney and spleen. Histopathology showed reduced inflammation in all target tissues, especially in the heart. Interestingly unlike TGF-? (-/-) mice, the TGF-?1(-/-/TG) mice have glomerulonephritis in their kidneys similar to the TG mice. Thus, the phenotype of TGF-?1 (-/-/TG) animal model indicates the potential role of circulating active-TGF-_1 in reducing inflammation, but its failure to rescue lethality in TGF-_1 null mice indicates a critical role of autocrine TGF-?1. The studies were also carried out to develop animal model to characterize the role of the TGF-? signaling pathway in squamous cell carcinoma (SCC) and test it for the cytokine receptor-directed cytotoxin therapy. Mice with the TGF-? receptor RI locus flanked with loxP sites were crossed with the neurofilament-H (NF-H) Cre mouse line 7 to generate TGF-? RI conditional knockout (COKO) mice. Tumors from the COKO mice were processed either for histological analysis or for establishing primary cell cultures using standard procedures. Recombinant IL-13 receptor-directed cytotoxin was expressed in E. coli and the purified material was used for cytotoxicity assays on SCC primary cell cultures. Twenty percent of the 6-month old COKO mice developed tumors located in either the head and neck or the perianal region. The tumor burden increased with age. Tumors were typical SCC: their cells stained positive for keratinocyte markers and were surrounded by unusually large clusters of Merkel cells. Similar to human SCC tumor cell lines, some SCC primary cultures derived from COKO mice were highly sensitive to the cytotoxic effects of IL-13 receptor- directed cytotoxin. The targeted deletion of TGF-? signaling in the neuronal cells populating both the central and peripheral nervous system led to SCC tumors in the head and neck, and the perianal regions. These regions are rich in Merkel cells and their abnormal clustering around the tumors indicate their potential involvement in SSC in COKO mice. Approximately 30% of human SCC tumors are known to express high levels of IL-13 receptors and are highly sensitive to IL-13 cytotoxin. Since SCC cells derived from COKO mice exhibit similar unique characteristics, these mice may be a suitable animal model for studying receptor-directed cytotoxin therapy.

细胞因子和生长因子在免疫功能的正常内稳态中起着关键作用。转化生长因子-β1是炎症反应的主要调节因子。我们早先建立了转化生长因子-1-/-小鼠，表现出与白细胞与内皮细胞黏附增加相关的多灶性炎症，MHC-I和-II类抗原的异常表达，以及类似于干燥综合征的自身免疫表现。此外，去除CD8细胞改善了转化生长因子-1基因缺失小鼠的健康状况。为了阐明转化生长因子-1在MHC异常表达中的作用，在MHC-I和MHC-II缺乏的背景下产生了转化生长因子-1缺失的小鼠。转化生长因子-1X MHC-I缺失小鼠的寿命延长，炎症减轻，自身免疫表现减少，骨髓生成显著增加。尽管转化生长因子-1X MHC-II小鼠缺乏炎症和自身免疫反应，但它们的新生儿死亡率与增加的骨髓生成和化生有关。这些结果表明，转化生长因子-β1在白细胞成熟和功能中起主导作用。为了评估循环中活性转化生长因子-1水平的治疗潜力，我们通过将转化生长因子-1(+/-)小鼠与转基因小鼠(TG)杂交，在转化生长因子-1缺失背景(转化生长因子-1(-/-/TG)的背景下产生内分泌表达活性转化生长因子-1的小鼠(-/-/TG)，转基因小鼠(TG)在肝脏中特异性表达重组转化生长因子-1并在血液中分泌。转化生长因子-1(-/-/TG)小鼠的存活曲线与转化生长因子-1(-/-)小鼠相似，表明未能挽救致死表型。然而，转化生长因子-β1(-/-/TG)组小鼠的血清转化生长因子-β1水平恢复到正常水平，在所有组织中都有表达，尤其是在肾脏和脾组织。组织病理学显示，所有靶组织的炎症反应都有所减轻，特别是心脏。有趣的是，与转化生长因子β(-/-)小鼠不同的是，转化生长因子-1(-/-/甘油三酯)小鼠的肾脏中的肾小球肾炎与转基因小鼠相似。因此，动物模型的表型提示了循环活性的转化生长因子-1在抗炎中的潜在作用，但它不能挽救转化生长因子-1缺失的小鼠的致死性，表明了自分泌转化生长因子-1的关键作用。在鳞状细胞癌(SCC)中的信号通路，并检测其用于细胞因子受体导向的细胞毒素治疗。有转化生长因子的小鼠？将带有loxP位点的受体RI基因座与神经丝蛋白-H(Nf-H)Cre小鼠品系7杂交，产生转化生长因子？RI条件性基因敲除(Coko)小鼠。来自可可小鼠的肿瘤被处理以进行组织学分析或使用标准程序建立原代细胞培养。重组IL-13受体导向的细胞毒素在大肠杆菌中表达，并用于原代培养的SCC细胞毒活性测定。在6个月大的可可老鼠中，有20%的小鼠患上了位于头部和颈部或肛周的肿瘤。肿瘤负担随年龄增长而增加。肿瘤是典型的鳞状细胞癌：它们的细胞角质形成细胞标记物染色阳性，周围环绕着异常大的默克尔细胞簇。与人鳞状细胞癌肿瘤细胞系相似，Coko小鼠来源的一些原代培养的SCC细胞对IL-13受体导向的细胞毒素的细胞毒作用高度敏感。靶向缺失转化生长因子-？中枢和外周神经系统中的神经细胞中的信号传递导致了头颈部和肛周区域的鳞癌肿瘤。这些区域富含Merkel细胞，它们在肿瘤周围的异常聚集表明它们可能参与了Coko小鼠的SSC。大约30%的人鳞状细胞癌肿瘤表达高水平的IL-13受体，并对IL-13细胞毒素高度敏感。由于Coko小鼠来源的SCC细胞具有相似的独特特性，这些小鼠可能是研究受体导向细胞毒素治疗的合适的动物模型。