Leukocyte subset identification by single cell analysis of BTB-ZF gene

描述(由申请人提供)：已发现数量较少的淋巴细胞亚群具有强大的免疫调节功能。事实上，T细胞亚群的识别，如调节性T细胞、T滤泡辅助细胞、自然杀伤T细胞和产生IL-17的T细胞 在理解获得性免疫反应方面，细胞一直是最重要的进展之一。BTB-锌指(BTB-ZF)转录因子由N-末端蛋白-蛋白质相互作用结构域(BTB/POZ)和C-末端锌指结构域共同定义。BTB-ZF家族中的几个成员已经成为控制免疫系统特定细胞类型的发育或功能的基本、非冗余因素。例如，BTB-ZF基因已被证明控制T细胞对B细胞的承诺(LRF)，CD4对CD8谱系的承诺(ThPOK)，胸腺细胞对先天T细胞谱系的承诺(PLZF)，T滤泡辅助T细胞(Bcl6)的发育以及B细胞向生发中心B细胞的分化(Bcl6)。这个基因家族的其他成员显然会影响免疫反应，但影响方式不太明确。在R21的应用中，我们建议探索BTB-ZF基因家族的表达，以定义新的白细胞亚群效应群体。我们将开发一套基于BAC的转基因，在BTB-ZF基因的调控元件的控制下同时表达GFP和Cre。初步研究表明，每个选定的BTB-ZF基因都在免疫系统中表达，在发育过程中受到调节，并有可能控制一组新细胞的功能。我们已做好准备进行这些研究，并已证明BAC记者系统是一种非常有效的方法。事实上，我们强烈认为，这种单细胞表达模型对于识别白细胞亚群和研究这一重要的新转录因子家族是必不可少的。到目前为止已经确定的BTB-ZF基因的戏剧性和非冗余功能清楚地支持了我们建议在R21应用的背景下开发和评估的试剂的重要性。我们相信，这一努力将极大地促进对免疫反应转录控制的理解。最后，由于这个转录因子家族在人类中高度保守，我们的研究有可能导致新的诊断测试和/或治疗。

DESCRIPTION (provided by applicant): Numerically minor subsets of lymphocytes have been found to have potent immunomodulatory functions. Indeed, the identification of T cell subsets such as regulatory T cells, T follicular helper cells, natural killer T cells and IL-17 producing T cell have, cumulatively, represented one of the most important advances in understanding the adaptive immune response. BTB-zinc finger (BTB-ZF) transcription factors are defined by an N-terminal protein-protein interaction domain (BTB/POZ) domain, coupled with a C-terminal zinc finger domain. Several members of this BTB-ZF family have emerged as fundamental, non-redundant factors that control the development or function of specific cell types of the immune system. For example, BTB-ZF genes have been shown to control T cell versus B cell commitment (LRF), CD4 versus CD8 lineage commitment (ThPOK), the commitment of thymocytes to innate T cell lineages (PLZF), the development of T follicular helper T cells (Bcl6) as well as differentiation of B cells into germinal center B cells (Bcl6). Additional members of ths gene family clearly influence the immune response, but in less well defined ways. In this R21 application we propose to explore BTB-ZF gene family expression to define novel leukocyte subset effector populations. We will develop a set of BAC-based transgenes that express both GFP and Cre under the control of the regulatory elements for BTB-ZF genes. Preliminary studies suggest that each of the selected BTB-ZF genes is expressed in the immune system, is regulated during development and, potentially, controls the function of a novel subset of cells. We are well positioned to carry out these studies and have already proven that the BAC reporter system is a highly effective approach. Indeed, we would strongly argue that this single cell expression model is essential for identifying leukocyte subsets and for the study of this important new family of transcription factors. The dramatic and nonredundant functions of the BTB-ZF genes that have thus far been identified clearly support the significance of the reagents we propose to develop and evaluate in the context of this R21 application. We believe that this effort will significantly advance the understanding of the transcriptional control of immune responses. Finally, since this family of transcription factors is highly conserved in humans, our studies have the potential to lead to new diagnostic tests and/or therapies.