Autoantigen Arrays to Select Vaccines for Model of PBC

描述(由申请人提供)： 这项建议的广泛和长期目标是使用自身抗体图谱来指导选择DNA耐受疫苗来治疗被称为实验性自身免疫性胆管炎(EAC)的原发性胆汁性肝硬变(PBC)动物模型。我们将测试这一假设，即大规模的、平行的自身抗体图谱检测可以用于探索表位扩散、炎性细胞因子子集在自身免疫的启动和传播中所起的作用，以及最终在选择抗原特异性耐受疗法中所起的作用。我们将使用生化、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，以探索这项提案的三个具体目标：(I)构建适合于鉴定人和小鼠自身抗体的PBC自身抗原微阵列。该阵列将用于识别EAC模型中的主要B细胞表位，并确定EAC中表位扩散的精确顺序。(Ii)以确定EAC的发生发展是否需要白介素4(IL-4)和干扰素-γ(IFNy)。Th1/Th2细胞失衡被认为在某些自身免疫性疾病中起重要作用，而IL-4和IFN3‘是自身免疫发病机制中的关键细胞因子。(三)应用DNA疫苗通过编码自身抗原的cDNA预防和治疗EAC，该cDNA是利用第一个目标开发的蛋白质微阵列鉴定的。这项建议的结果将决定新描述的PBC EAC动物模型的长期有用性。使用编码显著自身抗原的质粒成功治疗EAC可能预示着人类定制化、抗原或组织特异性耐受治疗的时代的到来。

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.