dnTGF Beta RII Mice and PBC

dnTGF Beta RII 小鼠和 PBC

• 批准号: 8152134
• 负责人: MERRILL E GERSHWIN
• 金额: $ 43.34万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 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/8152134
• 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

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（由申请人提供）：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，其特征是门户道淋巴细胞浸润，胆道上皮细胞和抗降节性抗体抗体（AMAS）的选择性破坏。我们的实验室在进行PBC的研究方面发挥了重要作用，该研究始于大约25年前的识别和克隆线粒体自动抗原。但是，PBC的突破需要动物模型，在过去的几年中，我们的实验室描述了几种此类模型。但是，在更大相似性中概括的模型人类PBC的特征是一种小鼠转基因，用于在CD4启动子的方向下指示TGF2受体II型（DNTGF2-RII）的显性负形式的表达。使用导致该模型发展的R21奖项的资金，我们报告说，DNTGF2-RII小鼠具有与人类相同特异性的AMA，在血清和肝脏中都具有相似的细胞因子谱，并且具有与人类类似于人类的肝组织学和免疫组织化学。此外，脾脏DNTGF2-RII CD8+ T细胞转移到B6 rag1 - / - 小鼠中会转移自身免疫性胆管炎，而dntGF-2RII CD4+ T细胞的相似转移也会转移结肠炎。我们建议利用这些和其他数据，这是我们R21奖的逻辑扩展，以广泛剖析导致胆道自身免疫性疾病的免疫发病机制。特别是，我们提出了三个关键目标。 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.其次，我们将通过利用我们在I类四级技术领域的专业知识以及使用受限的TCR库库来定义DNTGF2-RII CD8+ T细胞自身免疫性是否特定于抗原。最后，我们最近发表了在DNTGF2-RII小鼠中删除IL-12P40而不是IFN-3的删除，从而阻止了疾病的发展。鉴于在人类PBC的全基因组关联研究中证明了最近的IL-12多态性，这令人兴奋。我们将利用生产独特的鼠构建体的能力，并将剖析产生疾病的细胞因子介导的途径。 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）小鼠到DNTGF-2RII小鼠。此外，由于DNTGF-2RII小鼠的血清IL-22显着升高，因此我们还将将IL-22 - / - 小鼠回到DNTGF-2RII小鼠上，以进一步定义IL-22的保护或炎症作用。我们认为，使用我们独特的鼠构建体以及对自身免疫性胆管炎机制的严格解剖，将为成功调节人类PBC提供重要的线索。 公共卫生相关性：原发性胆汁肝硬化是一种人类的神秘自身免疫性疾病，其特征是自身抗体和肝脏胆道系统的炎症。缺乏动物模型，这种疾病的进展受到了阻碍。我们的实验室建议利用一个概括人类PBC的许多特征的模型，并研究疾病的机制，认为这种数据将导致人类PBC的突破。