Multiplex Autoantibody Profiling in SLE

描述(由申请人提供)：这项提案的广泛、长期目标是开发、验证和使用蛋白质和多肽自身抗原微阵列来分析生物液中发现的自身抗体。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。我们已经减少了在许多人类自身免疫性疾病中使用大规模阵列来识别自身抗体图谱的做法。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，以探索这项提案的五个具体目标：(I)确定打印蛋白质、多肽和核糖核蛋白复合体的通用表面化学；使用高度特征化的血清样本和单抗验证自身抗体与个体特征的结合；(Iii)构建和验证用于检测自身抗体的全面CTD自身抗原阵列；使用自身抗原微阵列鉴定自发和可诱导的系统性红斑狼疮小鼠模型的血清和组织来源的自身抗体；为了验证自身抗体谱可用作抗原特异的、基于DNA质粒的耐受疫苗治疗的动物耐受性的替代标记这一假设。这一提议的结果可能会阐明B淋巴细胞及其分泌产物在自身免疫中的作用扩大，或许预示着人类定制化、抗原或组织特异性耐受治疗的时代即将到来。此外，蛋白质微阵列技术的进一步发展将在免疫学、功能基因组学和蛋白质组学领域具有广泛的应用前景。

DESCRIPTION (provided by applicant): The broad, long-term objective of this proposal is to develop, validate, and employ protein and peptide autoantigen microarrays for profiling autoantibodies found in biological fluids. 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 have reduced to practice the use of large-scale arrays to identify autoantibody profiles in many human autoimmune diseases. We will use biochemical, immunological, and molecular biological techniques to validate and extend our ongoing protein array platform in exploring five specific aims in this proposal: (i.) to identify a universal surface chemistry for printing protein, peptide, and ribonucleoprotein complexes; (ii.) to validate autoantibody binding to individual features using highly-characterized serum samples and monoclonal antibodies; (iii.) to construct and validate a comprehensive CTD autoantigen array for detection of autoantibodies; (iv.) to use autoantigen microarrays to characterize serum and tissue-derived autoantibodies from spontaneous and inducible murine models of SLE; and (v.) to test the hypothesis that autoantibody profiling can be used as a surrogate marker of tolerance in animals treated with antigen-specific, DNA plasmid-based, tolerizing vaccines. The results of this proposal may elucidate an expanded role for B lymphocytes and their secreted products in autoimmunity, perhaps heralding an era of customized, antigen- or tissue- specific tolerizing therapy in humans. Moreover, further development of protein microarray technology will have broad applications to the fields of immunology, functional genomics, and proteomics.