Role of CD8+ T Cells in Innate Immune Responses

描述（由申请方提供）：本提案旨在了解在细菌感染期间在不存在同源抗原的情况下应答的CD 8 + T细胞的发育、快速功能和定位，假设CD 8 + T细胞是IFN-γ早期先天产生的重要贡献者。具体目标是：我：分析记忆性CD 8 + T细胞和NK细胞在单核细胞增生李斯特菌（LM）感染中的存活、定位和保护能力。在这个目标中，我们将解剖NK和记忆CD 8 + T细胞之间的保护能力的差异，以非抗原特异性的方式响应LM。我们将使用免疫细胞化学来分析LM感染靶器官内转移细胞的应答群体的位置。此外，将使用免疫细胞化学和流式细胞术测量记忆性CD 8 + T和NK细胞在响应LM后存活的能力。II：确定CD 8 + T细胞在辅助CD 4 + T细胞极化中的作用。我们的数据显示，记忆性CD 8 + T细胞快速分泌IFN-γ，不依赖于同源抗原，因此在先天免疫应答中发挥重要作用，这在通过分泌细胞因子促进TH 1发育中至关重要。我们现在将直接测试CD 8 + T细胞和NK细胞通过使用共转移方案分泌IFN-γ来影响TH 1发育的能力，以分析响应病原体的CD 4 + T细胞的极化。III：分析IL-12和IL-18对初始和效应/记忆CD 8 + T细胞的作用。在这个目标的第一部分，我们将确定哪些细胞因子，如果有的话，是必需的，以建立IL-12/IL-18的反应过程中的幼稚CD 8 + T细胞转化为引发，效应细胞。在目标的第二部分中，我们将确定通过IL-12和IL-18受体的信号传导的功能结果，并使用微阵列分析和实时RT-PCR将其与通过TCR的信号传导进行比较。这些研究将进一步加深我们对CD 8 + T细胞介导的病原体先天免疫反应的理解，并使我们能够更好地对抗传染病。

DESCRIPTION (provided by applicant): This proposal is aimed at understanding the development, rapid function, and localization of CD8+ T cells that respond in the absence of cognate antigen during bacterial infections, with the hypothesis that CD8+ T cells are important contributors to the early, innate production of IFN-gamma. The specific aims are: I: To analyze the survival, localization and protective ability of memory CD8+ T cells vs. NK cells during a Listeria monocytogenes (LM) infection. In this aim we will dissect the differences in protective ability between NK and memory CD8+ T cells responding to LM in a non-antigen specific fashion. We will use immunocytochemistry to analyze the location of the responding populations of transferred cells within the organs targeted by LM infection. In addition, the ability of memory CD8+ T and NK cells to survive after responding to LM will be measured using immunocytochemistry and flow cytometry. II: To determine the role of CD8+ T cells in aiding in the polarization of CD4+ T cells. Our data shows that memory CD8+ T cells rapidly secrete IFN-gamma independent of cognate antigen and thus play an important role in the innate immune response, which is essential in promoting TH1 development through the secretion of cytokines. We will now directly test the ability of CD8+ T cells and NK cells to influence TH1 development by secreting IFN-gamma using co-transfer protocols to analyze the polarization of CD4+ T cells in response to pathogens. III: To analyze the effects of IL-12 and IL-18 on naive and effector/memory CD8+ T cells. In the first part of this aim, we will determine which cytokines, if any, are required in order to establish IL-12/IL-18 responsiveness during the transition of naive CD8+ T cells into primed, effector cells. In the second part of the aim, we will determine the functional outcomes of signaling through the IL-12 and IL-18 receptors and compare that with signaling through the TCR using microarray analysis and real-time RT-PCR. These studies will further our understanding of innate immune responses to pathogens mediated by CD8+ T cells and allow us to better combat infectious diseases.