Multiplex Autoantibody Profiling in SLE

描述（由申请人提供）：本提案的广泛，长期目标是开发，验证和使用蛋白质和肽自身抗原微阵列来分析生物液体中发现的自身抗体。我们将验证这样一个假设，即大规模、平行检测自身抗体谱可用于探索表位扩散、炎症细胞因子子集在自身免疫的启动和传播中所起的作用，并最终选择抗原特异性耐受疗法。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鉴定自身抗体谱的实践。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，以探索本提案中的五个具体目标：(i)确定用于打印蛋白质，肽和核糖核蛋白复合物的通用表面化学；（ii）使用高度表征的血清样本和单克隆抗体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综合CTD自身抗原阵列；（iv）使用自身抗原微阵列来表征自发和诱导型SLE小鼠模型的血清和组织来源的自身抗体；(v)验证假设，即自身抗体谱分析可作为抗原特异性、基于DNA质粒的耐受性疫苗治疗动物的耐受性的替代标记物。这一建议的结果可能阐明了B淋巴细胞及其分泌产物在自身免疫中的扩展作用，可能预示着人类定制抗原或组织特异性耐受性治疗的时代。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等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.