Autoantigen Arrays to Select Vaccines for Model of PBC

描述（由申请人提供）： 该提案的广泛的长期目标是使用自身抗体谱来指导选择DNA耐受性疫苗，用于治疗称为实验性自身免疫性胆管炎（EAC）的原发性胆汁性肝硬化（PBC）动物模型。我们将测试的假设，大规模，平行检测自身抗体谱可用于探索表位扩散，炎症细胞因子的子集在自身免疫的启动和传播中发挥的作用，并最终在选择抗原特异性耐受治疗。我们将使用生物化学，免疫学和分子生物学技术来验证和扩展我们正在进行的蛋白质阵列平台，以探索本提案中的三个具体目标：（i）构建适合于人和鼠自身抗体鉴定的PBC自身抗原微阵列。该阵列将用于鉴定EAC模型中的显性B细胞表位，并确定EAC中表位扩展的精确顺序。（二）以确定白细胞介素-4（IL-4）和干扰素-γ（IFN γ）是否是EAC发展所必需的。Th 1和Th 2淋巴细胞之间的失衡被认为在一些自身免疫性疾病中起重要作用，并且IL-4和IFN 3 '代表自身免疫发病机制中的关键细胞因子。（三）使用DNA疫苗接种来预防和治疗EAC，所述DNA疫苗接种使用编码自身抗原的cDNA，所述自身抗原使用在第一个目的中开发的蛋白质微阵列鉴定。该建议的结果将确定新描述的PBC EAC动物模型的长期有用性。使用编码突出的自身抗原的质粒成功治疗EAC可能预示着人类定制的抗原或组织特异性耐受治疗的时代。

DESCRIPTION (provided by applicant): The broad, long-term objective of this proposal is to use autoantibody profiling to guide the selection of DNA tolerizing vaccines for the treatment of an animal model of primary biliary cirrhosis (PBC) called experimental autoimmune cholangitis (EAC). We will test the hypothesis that large-scale, parallel detection of autoantibody profiles can be used to explore epitope spreading, the role played by a subset of inflammatory cytokines in the initiation and propagation of autoimmunity, and ultimately in selection of antigen-specific tolerizing therapies. We will use biochemical, immunological, and molecular biological techniques to validate and extend our ongoing protein array platform in exploring three specific aims in this proposal: (i.) to construct a PBC autoantigen microarray suitable for the identification of human and murine autoantibodies. This array will be used to identify dominant B cell epitopes in the EAC model, and to determine the precise ordering of epitope spreading in EAC. (ii.) to determine whether interleukin-4 (IL-4) and interferon-gamma (IFNy) are required for the development of EAC. An imbalance between Thl and Th2 lymphocytes is thought to play an important role in some autoimmune diseases, and IL-4 and IFN3' represent critical cytokines in the pathogenesis of autoimmunity. (iii.) to employ DNA vaccination to prevent and treat EAC using cDNAs encoding autoantigens identified using protein microarrays developed in the first aim. The results of this proposal will determine the long-term usefulness of the newly described EAC animal model of PBC. Successful treatment of EAC using plasmids encoding prominent autoantigens may herald an era of customized, antigen- or tissue- specific tolerizing therapy in humans.