dnTGF Beta RII Mice and PBC

dnTGF Beta RII 小鼠和 PBC

基本信息

批准号：
8287116
负责人：
MERRILL E GERSHWIN
金额：
$ 42.51万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-30 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8287116
关键词：
Address Adoptive Transfer Animal Model Antibodies Antigens Autoantibodies Autoantigens Autoimmune Diseases Autoimmune Process Autoimmunity Backcrossings Belief Biliary Bone Marrow Bystander Effect CD4 Positive T Lymphocytes CD8B1 gene Cells Cholangitis Cloning Colitis Data Development Disease Dissection Dominant-Negative Mutation Effector Cell Environment Epithelial Cells Exploratory/Developmental Grant for Diagnostic Cancer Imaging Floods Funding Genetic Polymorphism Hepatic Histology Human Immune Immunohistochemistry In Vitro Infiltration Inflammation Inflammatory Interferons Interleukin-12 Interleukin-17 Laboratories Lead Light Liver Lymphocyte Mediating Memory Mitochondria Modeling Mus Ovum Pathogenesis Pathogenicity Pathology Pathway interactions Population Primary biliary cirrhosis Publishing Rag1 Mouse Relative (related person)Reporting Role Serum Signal Pathway Signal Transduction Specificity T cell response T memory cell T-Lymphocyte Technology Time Transforming Growth Factors Transgenes Transgenic Mice Work autoreactivity base biliary tract cytokine disease mechanisms study genome wide association study in vivo interleukin-12 subunit p40 interleukin-22 interleukin-23 model development novel prevent promoter public health relevance pyruvate dehydrogenase complex E2 receptor

项目摘要

DESCRIPTION (provided by applicant): Primary biliary cirrhosis (PBC) is a liver specific autoimmune disease characterized by portal tract lymphocytic infiltration, selective destruction of biliary epithelial cells and antimitochondrial antibodies (AMAs). Our lab has been instrumental in advancing the study of PBC, work which began nearly 25 years ago with the identification and cloning of the mitochondrial autoantigens. However, a breakthrough in PBC requires an animal model and over the past several years our laboratory has described several such models. However, the model which recapitulates in greater similarity the features of human PBC, is a mouse transgenic for directed expression of a dominant negative form of TGF2 receptor type II (dnTGF2-RII) under the direction of the CD4 promoter. Using funds from an R21 award that led to the development of this model, we have reported that dnTGF2-RII mice develop AMAs of the identical specificity as humans, similar cytokine profiles in both sera and liver and have liver histology and immunohistochemistry similar to humans with PBC. Further, adoptive transfer of splenic dnTGF2-RII CD8+ T cells into B6 Rag1-/- mice transfers autoimmune cholangitis, whereas similar transfer of dnTGF-2RII CD4+ T cells transfers colitis. We propose to take advantage of these and other data, a logical extension of our R21 award to extensively dissect the immunopathogenic mechanisms that lead to biliary autoimmune disease. In particular, we propose three critical aims. Firstly, we have demonstrated that dnTGF2-RII CD8+ T cells are sufficient to transfer biliary pathology to Rag1-/- recipients and we propose to define the mechanism of dnTGF2-RII CD8+ T cell mediated disease using transfer and mixed bone marrow chimeric studies and an ontogenetic analysis of CD8+ T cells and an analysis of CD8+ T cell cytokine expression. Secondly, we will define whether dnTGF2-RII CD8+ T cell autoimmunity is antigen specific by taking advantage of our expertise in class I tetramer technology and by using dnTGF2-RII mice with a restricted TCR repertoire. Finally, we have recently published that deleting IL-12p40, but not IFN-3, in dnTGF2- RII mice prevents the development of disease. This is exciting in light of recent IL-12 polymorphism demonstrated in genome-wide association studies of human PBC. We will take advantage of our ability to produce unique murine constructs and we will dissect the cytokine mediated pathways that produce disease. We will backcross p35-/- mice (lack IL-12), p19-/- mice (lack IL-23), p35/19-/- mice (lack IL-12 and IL-23), p35/40- /- mice (lack IL-12, IL-23, and homodimeric IL-12p40), or p35/IL-17A-/- mice (lack IL-12 and IL-17A) mice to dnTGF-2RII mice. Furthermore, as sera IL-22 is significantly elevated in dnTGF-2RII mice, we will also backcross IL-22-/- mice onto dnTGF-2RII mice to further define the protective or inflammatory role of IL-22. We submit that use of our unique murine constructs and our rigorous dissection of the mechanism of autoimmune cholangitis in this model will provide important clues towards successful modulation of human PBC. PUBLIC HEALTH RELEVANCE: Primary biliary cirrhosis is a cryptic autoimmune disease of humans characterized by autoantibodies and inflammation of the biliary system of the liver. Progress in this disease has been hampered by the absence of an animal model. Our lab proposes to take advantage of a model that recapitulates many of the features of human PBC and study the mechanisms of disease with the belief that such data will lead to a breakthrough in human PBC.

描述（申请人提供）：原发性胆汁性肝硬化（PBC）是一种肝脏特异性自身免疫性疾病，其特征是汇管道淋巴细胞浸润、胆管上皮细胞和抗线粒体抗体（AMA）的选择性破坏。我们的实验室在推进 PBC 研究方面发挥了重要作用，这项工作始于近 25 年前，鉴定和克隆了线粒体自身抗原。然而，PBC 的突破需要动物模型，在过去的几年里，我们的实验室已经描述了几个这样的模型。然而，与人类 PBC 特征更加相似的模型是转基因小鼠，在 CD4 启动子的指导下定向表达显性失活形式的 II 型 TGF2 受体 (dnTGF2-RII)。利用导致该模型开发的 R21 奖项的资金，我们报道了 dnTGF2-RII 小鼠产生了与人类相同特异性的 AMA，血清和肝脏中具有相似的细胞因子谱，并且具有与人类相似的肝脏组织学和免疫组织化学。中国人民银行。此外，将脾脏 dnTGF2-RII CD8+ T 细胞过继转移到 B6 Rag1-/- 小鼠中会转移自身免疫性胆管炎，而类似的 dnTGF-2RII CD4+ T 细胞转移也会转移结肠炎。我们建议利用这些数据和其他数据，这是我们 R21 奖项的逻辑延伸，广泛剖析导致胆道自身免疫性疾病的免疫致病机制。我们特别提出三个关键目标。首先，我们已经证明 dnTGF2-RII CD8+ T 细胞足以将胆道病理转移至 Rag1-/- 受体，我们建议使用转移和混合骨髓嵌合研究和混合研究来定义 dnTGF2-RII CD8+ T 细胞介导的疾病的机制。 CD8+ T 细胞的个体发育分析和 CD8+ T 细胞细胞因子表达的分析。其次，我们将利用我们在 I 类四聚体技术方面的专业知识并使用具有受限 TCR 库的 dnTGF2-RII 小鼠来定义 dnTGF2-RII CD8+ T 细胞自身免疫是否具有抗原特异性。最后，我们最近发表了在 dnTGF2-RII 小鼠中删除 IL-12p40 而不是 IFN-3 可以预防疾病的发展。鉴于最近在人类 PBC 全基因组关联研究中证明的 IL-12 多态性，这是令人兴奋的。我们将利用我们生产独特小鼠结构的能力，我们将剖析产生疾病的细胞因子介导的途径。我们将回交 p35-/- 小鼠（缺乏 IL-12）、p19-/- 小鼠（缺乏 IL-23）、p35/19-/- 小鼠（缺乏 IL-12 和 IL-23）、p35/40-/ - 小鼠（缺乏 IL-12、IL-23 和同二聚体 IL-12p40）或 p35/IL-17A-/- 小鼠（缺乏 IL-12）和 IL-17A) 小鼠至 dnTGF-2RII 小鼠。此外，由于 dnTGF-2RII 小鼠的血清 IL-22 显着升高，我们还将 IL-22-/- 小鼠与 dnTGF-2RII 小鼠回交，以进一步确定 IL-22 的保护或炎症作用。我们认为，在该模型中使用我们独特的小鼠构建体以及我们对自身免疫性胆管炎机制的严格剖析将为成功调节人类 PBC 提供重要线索。公共卫生相关性：原发性胆汁性肝硬化是一种隐秘的人类自身免疫性疾病，其特征是自身抗体和肝脏胆道系统炎症。由于缺乏动物模型，这种疾病的进展受到阻碍。我们的实验室建议利用一个概括人类原发性胆汁性胆管炎的许多特征的模型来研究疾病的机制，相信这些数据将为人类原发性胆汁性胆管炎带来突破。