Regulation of effector T cell function by Blimp-1 in a murine model of colitis

描述（由申请方提供）：通过生理和病理生理炎症之间的微妙平衡维持肠粘膜的稳态，确保对病原微生物的有效反应，并避免对肠道植物群的高反应性。这种平衡的破坏导致慢性炎性病症的发展，例如溃疡性结肠炎和克罗恩病，统称为炎症性肠病（IBD）。T淋巴细胞是IBD的主要效应细胞，因此，干扰T细胞功能可能是治疗IBD的潜在策略。然而，IBD中不受控制的T细胞应答的确切机制尚未得到很好的理解。我们和其他人最近发现转录抑制因子B淋巴细胞诱导成熟蛋白-1（Blimp-1）是T细胞稳态和功能的重要调节因子。Blimp-1最初因其在分泌抗体的浆细胞的终末分化中的关键作用而为人所知。最近的研究表明，Blimp-1也在活化的T细胞中高度表达。具有Blimp-1的T细胞特异性缺失的小鼠在外周中积累“抗原经历的”细胞并自发地发展结肠炎，而不损害其他器官，这表明Blimp-1是肠粘膜处的适当T细胞功能所必需的。基于这些发现和以前在其他细胞谱系中的研究，这些研究确定了Blimp- 1控制几种遗传程序，我们假设在T细胞中Blimp-1调节遗传程序对于控制肠粘膜的反应性至关重要，并且在T细胞中Blimp-1的遗传靶点的鉴定应该揭示与粘膜稳态有关的新分子途径。在当前的提案中，我们将通过以下具体目标来测试这一假设：1）表征Blimp-1CKO小鼠中的结肠炎，以深入了解在不存在Blimp-1的情况下被破坏的效应机制;以及2）产生并比较Blimp-1充足和缺乏的致结肠炎T细胞，以鉴定并验证这些细胞中受Blimp-1调节的基因。为了实现这些目标，我们将使用我们以前产生的Blimp-1 CKO小鼠。首先，我们将进行表型和功能分析，以确定在这些小鼠中引起疾病的T细胞亚群。其次，我们将使用过继转移实验来产生Blimp-1充足和缺乏的致大肠杆菌性T细胞，用于基因表达研究（包括RNA微阵列），以鉴定由Blimp-1控制的分子途径。我们预计，在完成本文提出的实验后，我们将确定Blimp-1在T细胞中的新遗传靶点，并发现对调节肠粘膜中效应T细胞功能重要的新途径。对这些途径的进一步探索可以导致开发新的治疗方法来治疗IBD和潜在的其他炎症性疾病。公共卫生相关性：肠道中T细胞反应失衡导致胃肠道慢性炎症。我们正在寻求阐明调节机制，控制T细胞效应功能在肠粘膜。这些机制的鉴定将有助于开发新的治疗方法来治疗IBD。

DESCRIPTION (provided by applicant): Homeostasis in the intestinal mucosa is maintained by a delicate balance between physiological and pathophysiological inflammation that assures efficient response against pathogenic organisms and avoidance of hyperresponsiveness to the commensal flora. Disruption of this balance results in the development of chronic inflammatory conditions, such as ulcerative colitis and Crohn's disease, collectively termed Inflammatory Bowel Disease (IBD). T lymphocytes are major effector cells in IBD, thus, interfering with T cell function could be a potential strategy to treat IBD. However, the precise mechanisms underlying the uncontrolled T cell responses in IBD are not well understood. We and others have recently identified the transcriptional repressor B Lymphocyte Induced Maturation Protein-1 (Blimp-1) as an important regulator of T cell homeostasis and function. Blimp-1 was first known for its crucial role in the terminal differentiation of antibody-secreting plasma cells. Recent studies show that Blimp-1 is also highly expressed in activated T cells. Mice with a T cell-specific deletion of Blimp-1 accumulate "antigen-experienced" cells in the periphery and spontaneously develop colitis, without compromising other organs, indicating that Blimp-1 is required for proper T cell function at the intestinal mucosal. Based on these findings and previous studies in other cell lineages, which established that Blimp- 1 controls several genetic programs, we hypothesize that in T cells Blimp-1 regulates genetic programs crucially important to control responsiveness in the intestinal mucosa, and identification of genetic targets of Blimp-1 in T cells should reveal new molecular pathways implicated in mucosal homeostasis. In the current proposal we will test this hypothesis through the following specific aims: 1) characterize the colitis in the Blimp-1CKO mice to gain insight into the effector mechanisms disrupted in the absence of Blimp-1; and 2) generate and compare Blimp-1 sufficient and deficient colitogenic T cells to identify and validate genes regulated by Blimp-1 in these cells. To accomplish these aims we will use the Blimp-1 CKO mice we have previously generated. First, we will conduct phenotypical and functional analysis to identify the T cell subpopulations causing disease in these mice. Second, we will use adoptive transfer experiments to generate Blimp-1sufficient and deficient colitogenic T cells to be used in gene expression studies (including RNA microarray) to identify molecular pathways controlled by Blimp-1. We anticipate that upon completion of the experiments proposed here, we will have identified new genetic targets of Blimp-1 in T cells and uncovered new pathways important for the regulation of effector T cell function in the intestinal mucosal. Further exploration of these pathways can lead to the development of new therapeutic approaches to treat IBD and potentially other inflammatory disorders. PUBLIC HEALTH RELEVANCE: Unbalanced T cell responses in the intestines result in chronic inflammation of the GI tract. We are seeking to elucidate regulatory mechanisms that control T cell effector function in the intestinal mucosa. Identification of these mechanisms will contribute to development of new therapeutic approaches to treat IBD.