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剂。我们较早产生的TGF-？1 - / - 小鼠表现出与白细胞对内皮的粘附增加有关的多灶性炎症，MHC类-I类和-II抗原的异常表达以及与Sjogren综合征类似的自身免疫性表现。此外，CD8细胞的耗尽改善了TGF-？1个无效小鼠的健康状况。为了描述TGF-？1在异常MHC表达中的作用，在MHC-I和MHC-II缺陷背景中产生了TGF-？1 NULL小鼠。 TGF-？1 X MHC-I NULL小鼠的寿命增加与炎症减少，自身免疫性表现减少并显着升高骨髓病。尽管TGF-？1 X MHC-II小鼠表现出缺乏炎症和自身免疫反应，但它们显示出与骨髓病和化学症增加有关的新生儿死亡率。这些结果表明TGF-？1在白细胞成熟和功能中的主要作用。 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 血。 TGF-_1（ - / - /TG）小鼠表现出类似于TGF-？1（ - / - ）小鼠的生存谱，表明未能营救致命表型。但是，thetgf-？1（ - / - /tg）小鼠的血清TGF-？1水平恢复为正常水平，在所有组织中表达肾脏和脾脏中的所有组织。组织病理学显示所有靶组织，尤其是心脏的炎症降低。有趣的是，与TGF-不同？ （ - / - ）小鼠，TGF-？1（ - / - /TG）小鼠在类似于TG小鼠的肾脏中肾小球肾炎。因此，TGF-？1（ - / - /TG）动物模型的表型表明循环Active-TGF-_1在减少炎症中的潜在作用，但其未能挽救TGF-_1 NULL小鼠的致死性未能表明自动分泌TGF-？1的关键作用。还进行了研究以开发动物模型以表征TGF-的作用？鳞状细胞癌（SCC）中的信号通路，并测试细胞因子指导的细胞毒素治疗。带有TGF-的小鼠？与LOXP位点两侧的受体RI基因座与神经丝H（NF-H）CRE小鼠线7交叉，以生成TGF-？ RI条件敲除（COKO）小鼠。处理来自Coko小鼠的肿瘤进行组织学分析或使用标准程序建立原代细胞培养物。重组IL-13受体指导的细胞毒素在大肠杆菌中表达，并将纯化的材料用于SCC原代细胞培养物的细胞毒性测定。 6个月大的Coko小鼠中有20％在头部和颈部或植物区域出现肿瘤。肿瘤负担随着年龄的增长而增加。肿瘤是典型的SCC：它们的细胞对角质形成细胞标记呈阳性，并被异常大的默克尔细胞包围。与人类SCC肿瘤细胞系相似，一些源自COKO小鼠的SCC原发性培养物对IL-13受体定向的细胞毒素的细胞毒性作用高度敏感。 TGF-的目标删除？填充中枢和周围神经系统的神经元细胞中的信号传导导致头部和颈部的SCC肿瘤以及植物区域。这些区域富含默克尔细胞，肿瘤周围的异常聚类表明它们的潜在参与SSC在Coko小鼠中。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。由于源自COKO小鼠的SCC细胞具有相似的独特特征，因此这些小鼠可能是研究受体指导的细胞毒素治疗的合适动物模型。