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稳态中起关键作用。TGF-？1是炎症反应的主要调节因子。我们之前已经产生了TGF-？1-/-小鼠表现出与白细胞与内皮细胞粘附增加、MHC I类和II类抗原异常表达以及类似于舍格伦综合征的自身免疫表现相关的多灶性炎症。此外，耗尽的CD 8细胞改善健康状况的TGF-？1只无效小鼠。探讨TGF-？1异常MHC表达，TGF-？在MHC-I和MHC-II缺陷背景中产生1个无效小鼠。TGF-？IX MHC-I缺失小鼠表现出与炎症减少、自身免疫表现减少和骨髓生成显著升高相关的寿命增加。虽然TGF-？1X MHC-II小鼠表现出缺乏炎症和自身免疫应答，它们表现出与增加的骨髓生成和化生相关的新生儿死亡率。这些结果表明，TGF-？1在白细胞成熟和功能中的作用。为了评估治疗潜力的循环水平的活性TGF-？1，我们产生的小鼠内分泌表达活性TGF-？1个TGF-？1个空白背景（TGF-？1（-/-/TG））杂交。1（+/-）小鼠与转基因小鼠（TG）表达重组TGF-？1特别是在肝脏和分泌它在血液中。TGF-1（-/-/TG）小鼠表现出与TGF-？1（-/-）小鼠，表明未能挽救致死表型。血清TGF-？1水平的TGF-？1（-/-/TG）小鼠恢复到正常水平，在所有组织中表达，特别是在肾和脾中。组织学显示所有靶组织的炎症减少，特别是心脏。有趣的是，与TGF-？（-/-）小鼠，TGF-？1（-/-/TG）小鼠在其肾脏中具有与TG小鼠相似的肾小球肾炎。因此，表型的TGF-？1（-/-/TG）动物模型表明循环活性TGF-1在减轻炎症中的潜在作用，但其未能挽救TGF-1敲除小鼠的致死性，表明自分泌TGF-？1.研究还进行了开发动物模型，以表征TGF-？鳞状细胞癌（SCC）中的信号通路，并测试其用于细胞因子受体导向的细胞毒素治疗。小鼠与TGF-？受体RI位点两侧的loxP位点与神经毒-H（NF-H）Cre小鼠系7杂交，产生TGF-？RI条件性敲除（COKO）小鼠。使用标准程序处理来自COKO小鼠的肿瘤用于组织学分析或用于建立原代细胞培养物。重组IL-13受体介导的细胞毒素在大肠杆菌中表达。大肠杆菌，纯化的材料用于SCC原代细胞培养物的细胞毒性测定。20%的6个月大的COKO小鼠在头颈部或肛周区域发生肿瘤。肿瘤负荷随年龄增长而增加。肿瘤是典型的SCC：它们的细胞对角质形成细胞标记物染色呈阳性，并且被异常大的默克尔细胞簇包围。与人SCC肿瘤细胞系相似，一些来源于COKO小鼠的SCC原代培养物对IL-13受体导向的细胞毒素的细胞毒性作用高度敏感。TGF-？在中枢和周围神经系统的神经元细胞中的信号传导导致头颈部和肛周区域的SCC肿瘤。这些区域富含默克尔细胞，并且它们在肿瘤周围的异常聚集表明它们可能参与COKO小鼠中的SSC。已知大约30%的人SCC肿瘤表达高水平的IL-13受体，并且对IL-13细胞毒素高度敏感。由于来源于COKO小鼠的SCC细胞表现出相似的独特特征，这些小鼠可能是研究受体介导的细胞毒素治疗的合适动物模型。