Identification and characterization of molecules important in the immune system

描述（由申请人提供）：了解促进谱系定型和消除胸腺中自身反应性胸腺细胞的遗传机制与人类疾病有直接关系。我的研究涉及识别在胸腺的发育和分化中重要的新分子，并表征其在调节自身和非自身免疫反应中的分子基础。研究和理解自身耐受机制可能会导致对有害自身免疫性疾病患者的治疗。 我的实验设计涉及识别和表征适当的细胞发育和分化所必需的新分子。我使用基因组学（基因表达系列分析）， 鉴定在不同T细胞群中差异表达的特定基因产物。然后，我用一种新的RNAi方法来表征这些分子，以产生基因特异性缺陷小鼠。简言之，将含有基因特异性shRNA的慢病毒引入纯化的造血干细胞（HSC）中。然后用HSC重建RAG 2-/-受体小鼠以产生基因特异性敲低小鼠。 通过这些结合基因组和RNAi的方法，我鉴定并表征了MINK （畸形NIK相关激酶），这对胸腺中的阴性选择很重要。此外，我的目标是扩展这些方法的使用来表征SAGE识别的其他分子。 这项研究将有助于识别对免疫反应重要的新分子， 自我宽容此外，对其生物学功能的快速表征将有助于理解和找到治疗自身免疫性疾病的适当靶点。

DESCRIPTION (provided by applicant): Understanding the genetic mechanisms that promote lineage commitment and eliminate autoreactive thymocytes in the thymus has direct relation to human disease. My research involves identifying new molecules important in the development and differentiation of the thymus, and characterizing their molecular basis in regulating immune responses to both self and non-self. Studying and understanding self-tolerance mechanisms may result in therapeutic treatments for patients with harmful autoimmune diseases. My experimental design involves the identification and characterization of novel molecules necessary for proper cell development and differentiation. I use genomics (Serial Analysis of Gene Expression) to identify specific gene products differentially expressed in distinct T cell populations. I then characterize the molecules with a novel RNAi method to generate gene-specific deficient mice. In brief, lentivirus containing gene specific shRNA is introduced into purified hematopoetic stem cells (HSC's). RAG2 -/- recipient mice are then reconstituted with HSC's to generate gene-specific knockdown mice. With these combined genomic and RNAi-based methodologies I identified and characterized MINK (Misshapen-NIK-related kinase), which is important for negative selection in the thymus. Further, I aim to extend the use of these approaches to characterize other molecules identified by SAGE. This research will contribute to identifying new molecules important for immune responses and self-tolerance. Moreover, the rapid characterization of their biological functions will help to understand and find the proper targets for treating autoimmune diseases.