Novel genetic tools for the analysis of plasmacytoid dendritic cell function in vivo

摘要 浆细胞样树突状细胞(PDC)是一种独特的天然免疫细胞类型，能够迅速和 大量产生I型干扰素以应对病毒。PDC在这两种先天疾病中扮演着重要的角色 和对病毒感染的适应性免疫反应，而pDC的异常激活有助于 自身免疫性疾病，如系统性红斑狼疮。关于角色和分子的主要问题 PDC功能的基础仍未得到回答，这主要是由于缺少PDC特定的CRE删除器和 荧光报告菌株。该项目的总体目标是开发新的功能基因工具 对PDCs的分析。在目标1中，我们将使用候选的PDC特异性基因作为驱动来表达Cre 重组酶在pDC体内的表达。我们将研究Cre重组的特异性和有效性 产生的菌株，并将其用于体内的原则基因靶向验证。在目标2中，我们将使用相同的 一种生成PDC特异性荧光报告菌株并使用它显式显示PDC的方法 组织切片。总的来说，这些研究解决了模型系统研究PDC的主要未得到满足的需求 功能，因此将有助于体内研究先天免疫。

ABSTRACT Plasmacytoid dendritic cells (pDCs) represent a distinct innate immune cell type capable of rapid and massive production of type I interferon in response to viruses. pDCs play an important role in both innate and adaptive immune responses to viral infections, whereas aberrant activation of pDCs contributes to autoimmune diseases such as systemic lupus erythematosus. Major questions about the role and molecular basis of pDC function still remain unanswered, largely due to the absence of pDC-specific Cre deleter and fluorescent reporter strains. The overall goal of this project is to develop novel genetic tools for functional analysis of pDCs. In Aim 1, we will use a candidate pDC-specific gene as a driver to express Cre recombinase in pDCs in vivo. We will examine the specificity and efficiency of Cre recombination in the resulting strain and use it in a proof-of-principle gene targeting in vivo. In Aim 2, we will use the same approach to generate a pDC-specific fluorescent reporter strain and use it to explicitly visualize pDC in tissue sections. Collectively, these studies address a major unmet need for model systems to study pDC function, and therefore would facilitate in vivo studies of innate immunity.