Multiplex Autoantibody Profiling in SLE

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 用于分析在生物液中发现的自身抗体的自身抗原微阵列。我们将检验这一假设 即大规模、平行检测自身抗体谱可用于探索表位扩散、作用 炎性细胞因子在自身免疫的启动和传播中所起的作用 最终在抗原特异性耐受疗法的选择上。我们已经减少了使用大型- 用于识别许多人类自身免疫性疾病中的自身抗体图谱的规模阵列。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 阵列平台在本提案中探索五个具体目标：(I)确定一种普遍的表面化学 用于印刷蛋白质、多肽和核糖核蛋白复合体；验证自身抗体与 使用高度特征化的血清样本和单抗的个体特征；(3)建造 并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 用于鉴定自发和诱导小鼠血清和组织来源自身抗体的微阵列 狡猾的典范；和(V)为了检验自身抗体图谱可以用作替代标记的假设 用抗原特异性、DNA质粒基、耐受性疫苗治疗的动物的耐受性。结果是 这一建议可能会阐明B淋巴细胞及其分泌产物在 自身免疫，可能预示着定制的、抗原或组织特异性耐受治疗的时代即将到来 人类。此外，蛋白质微阵列技术的进一步发展将在以下方面有广泛的应用 免疫学、功能基因组学和蛋白质组学领域。

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 murinc models of SLY; 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-bascd, 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.