CD8 immunity to intracellular infection: Control by E-box transcription factors

描述(申请人提供)：CD8+T细胞对细胞内病原体如细菌、病毒和原生动物寄生虫的反应是宿主抵抗力的重要组成部分。这项建议利用细菌、单核细胞增生性李斯特菌和痘苗病毒的实验性传染病模型来解决宿主CD8+T细胞对潜在的生物恐怖主义微生物的免疫反应问题。 我们发现E蛋白转录因子及其抑制物Id2调节CD8+T细胞对细胞内病原体的反应，这是这些蛋白的一个新功能。我们的目标是在分子水平上了解这一转录调节家族如何影响CD8+T细胞从NAVE细胞到效应细胞再到记忆细胞的激活、增殖、分化和存活。虽然已知E蛋白在造血和淋巴细胞发育过程中调节许多关键的发育检查点、谱系承诺、增殖和生存，但这些重要蛋白在成熟T细胞中的功能尚不清楚。 我们假设在免疫反应中CD8+T细胞的激活和随后的记忆细胞的产生涉及到E蛋白转录靶标的调节。为了深入了解调控基因表达的E蛋白转录因子，在CD8+T细胞反应中受其活性调控的基因，以及抑制其活性如何调节记忆T细胞的形成，我们提出：目的1：确定在体内哪些E蛋白调节CD8+T细胞反应。我们将检测在感染单核细胞增多性李斯特氏菌和痘苗病毒期间，E2A、E2-2和Heb缺陷T细胞的免疫反应以及这些蛋白的DNA结合活性。目的：确定体内CD8+T细胞感染免疫应答过程中受E蛋白转录因子调控的分子途径。目的3：建立Id2报告小鼠品系，以确定Id2在免疫应答过程中的表达模式，并确定表达Id2的效应T细胞是否是记忆T细胞的前体。

DESCRIPTION (provided by applicant): The CD8+ T cell response to intracellular pathogens such as bacteria, viruses and protozoan parasites is an essential component of host resistance. This proposal utilizes experimental infectious disease models of bacteria, Listeria monocytongenes, and Vaccinia virus to address questions about the host CD8+ T cell immune response against potential microbial agents of bioterrorism. We have discovered that E protein transcription factors and their inhibitor, Id2, regulate the CD8+ T cell response to intracellular pathogens, which is a novel function for these proteins. It is our goal to understand at a molecular level how this family of transcriptional regulators influences the activation, proliferation, differentiation and survival of CD8+ T cells as they transition from naove to effector to memory cells. While the E proteins are known to regulate many key developmental check-points, lineage commitment, proliferation and survival during hematopoiesis and lymphocyte development, the function of these important proteins is unexplored in the mature T cell. We hypothesize that the activation of CD8+ T cells and subsequent generation of memory cells during the immune response involves the regulation of E protein- transcriptional targets. To gain insight into the specific E protein-transcription factors that regulate gene expression, the genes which are regulated by their activity during the CD8+ T cell response and how the inhibition of their activity regulates memory T cell formation, we propose to: Aim 1: Determine which E proteins regulate the in vivo CD8+ T cell response. We will examine the immune response by E2A, E2-2 and HEB-deficient T cells and the DNA-binding activity of each of these proteins during infection with Listeria monocytogenes and Vaccinia virus. Aim 2: Identify the molecular pathways controlled by E protein-transcription factors during the in vivo CD8+ T cell immune response to infection. Aim 3: Create an Id2-reporter mouse line to define the Id2 expression pattern during the immune response and determine if Id2 expressing effector T cells are the precursors to memory T cells.