Natural Killer Dendritic Cells in Listeria Infection

李斯特菌感染中的自然杀伤树突状细胞

• 批准号: 7232468
• 负责人: Ronald P Dematteo
• 金额: $ 22.16万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-01 至 2009-05-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7232468
• 关键词: Accounting; Antigens; Bioterrorism; Brefeldin A; Cells; Condition; Count; Cytolysis; Data; Dendritic Cells; Environment; Exposure to; Human; ITGAX gene; Immune response; Immunity; Immunologics; In Vitro; Infection; Inflammatory Response; Interferon Type II; Interferons; Interleukin-12; Interleukin-18; Label; Listeria; Listeria monocytogenes; Listeriosis; Liver; Lytic; Measures; Messenger RNA; Mus; Natural Killer Cells; Organ; Physiological; Polymerase Chain Reaction; Population; Production; Proteins; Research Personnel; Rodent; Role; Source; Spleen; Splenocyte; T-Lymphocyte; TLR9 gene; Techniques; Testing; Thymus Gland; Time; Wild Type Mouse; autocrine; base; cell type; cytokine; human mortality; in vivo; interferon alfacon-1; lymph nodes; novel; pathogen; research study; response

DESCRIPTION (provided by applicant): While there is general consensus that interferon-gamma (IFN-gamma) is vital to the clearance of Listeria monocytogenes, the cellular source of IFN-gamma has been controversial. Some investigators have concluded that NK cells are responsible, while others have implicated T cells. Recently, we have identified a novel cell type that secretes large amounts of IFN-gamma and exists in the spleen and other organs of normal mice. Because these unique cells lyse targets directly, activate naive T cells in vitro and in vivo, and express both the natural killer cell marker NK1.1 and the dendritic cell marker CD11c, we have labeled them as natural killer dendritic cells (NKDC). A few other investigators have recognized NKDC-like cells in rodents and humans. We have preliminary data that splenic NKDC produce IFN-gamma during Listeria infection in mice. Therefore, we hypothesize that NKDC are critical to the immune response against Listeria. While the lytic and antigen-presenting abilities of NKDC may be important, we have chosen to focus exclusively on their IFN-gamma production for this 2-year application. We propose to assess the biologic significance of IFN-gamma production by NKDC during Listeria infection in mice and dissect the mechanisms by which it is regulated. In Aim 1, we will determine the extent of NKDC involvement in IFN-gamma production during Listeria infection. To exclude the possibility that other cells make IFN-gamma in vivo, but are unable to do so once they are removed from their local environment in the spleen, we will make use of a recently described technique of in vivo intracellular cytokine analysis of splenocytes. In Aim 2, we will establish the mechanism of IFN-gamma production by NKDC during Listeria infection. Specifically, we will test IL-12, IL-18, and Toll-like receptor 9 as we have found these to regulate NKDC IFN-gamma production in response to bacterial CpG. In Aim 3, we will determine whether NKDC production of IFN-gamma is sufficient to induce an immune response against Listeria. We will adoptively transfer NKDC from wild-type mice into IFN-gamma deficient mice and then assess their response to Listeria infection. NKDC will be compared to NK cells and other splenic cell subsets that make IFN-gamma as identified in Aim 1. Our proposed experiments will begin to define the role of NKDC in the immune response to Listeria, a pathogen that is a cause of human mortality and a potential agent for bioterrorism, and provide a greater understanding of how this novel cell type may participate in other types of immunity.

描述（由申请人提供）：虽然有一般共识，即干扰素 - 伽马（IFN-GAMMA）对于清除单核细胞增生李斯特菌的清除至关重要，但IFN-GAMMA的细胞来源是有争议的。一些研究者得出的结论是，NK细胞是负责的，而另一些则牵涉到T细胞。最近，我们确定了一种新型的细胞类型，该细胞类型分泌了大量的IFN-gamma并存在于正常小鼠的脾脏和其他器官中。由于这些独特的细胞直接靶向靶向，在体外和体内激活幼稚的T细胞，并表达天然杀伤细胞标记物NK1.1和树突状细胞标记CD11C，因此我们将它们标记为天然杀手树突状细胞（NKDC）。其他一些研究人员已经认识到啮齿动物和人类中的NKDC样细胞。我们有初步数据，表明脾脏NKDC在小鼠李斯特菌感染期间产生IFN-gamma。因此，我们假设NKDC对于对李斯特菌的免疫反应至关重要。虽然NKDC的溶解和抗原呈现能力可能很重要，但我们选择专注于他们为期两年应用的IFN-gamma生产。我们建议评估小鼠李斯特菌感染期间NKDC产生IFN-gamma的生物学意义，并剖析调节其调节的机制。在AIM 1中，我们将确定NKDC在李斯特菌感染期间参与IFN-gamma产生的程度。为了排除其他细胞使IFN-gamma在体内产生的可能性，但一旦将它们从脾脏中的本地环境中删除，我们将利用一种最近描述的体内细胞内细胞因子细胞因子分析的脾细胞的技术。在AIM 2中，我们将在李斯特菌感染期间建立NKDC产生IFN-GAMMA的机制。具体而言，我们将测试IL-12，IL-18和类似Toll样受体9，因为我们发现它们以响应细菌CpG来调节NKDC IFN-gamma的产生。在AIM 3中，我们将确定IFN-GAMMA的NKDC产生是否足以诱导针对李斯特菌的免疫反应。我们将从野生型小鼠中传递NKDC转移到IFN-Gamma缺乏小鼠中，然后评估它们对李斯特菌感染的反应。 NKDC将与NK细胞和其他使IFN-GAMMA在AIM 1中确定的gamma进行比较。我们提出的实验将开始定义NKDC在对李斯特菌的免疫反应中的作用，这种病原体是人类死亡率的病原体，是人类死亡率的原因，并且对生物恐怖的潜在药物的一种潜在的药物，并且对这种新颖的细胞类型的影响更大，可以在其他类型的类型中参与其他细胞类型，并在其他类型的情况下参与其他类型。