Project 1 - Modulation of Tolerance and Autoimmunity by Inhibitory Receptors

项目摘要 PD 1和LAG 3在调节T细胞耐受和自身免疫中起关键作用。联合遗传缺陷 PD 1和LAG 3在小鼠中的协同作用导致致死性全身性自身免疫，证明了PD 1/LAG 3在小鼠中的协同作用。 宽容项目1的目标是确定PD 1和LAG 3控制Treg和自身免疫的机制。反应性CD 4 + FoxP 3- T细胞在耐受和自身免疫中的作用。这些都是基础和临床的问题 重要性，因为PD 1和LAG 3是T细胞耗竭的关键介质和有前途的治疗靶点， 在接受检查点治疗的癌症患者中， 封锁更深入地了解PD 1和LAG 3如何共同作用以介导耐受性， 有效的PD 1和LAG 3癌症治疗，同时最大限度地减少自身免疫。与EAE加重相比， Pdcd 1-/-小鼠，我们的初步数据显示Treg中缺乏PD 1的小鼠发生改善的EAE。相反地， Treg中的LAG 3缺失增加了EAE的严重程度，而Treg中的PD 1/LAG 3组合缺失减少了EAE。这些 研究结果强调需要确定PD 1/LAG 3信号如何整合在不同的细胞类型中， 了解干扰这些途径如何影响保护性Treg和致病性自身反应性CD 4 + FoxP 3- T 细胞基于这些数据，我们假设PD 1和LAG 3之间的协同作用是通过细胞和免疫系统来实现的。 Treg和CD 4 + FoxP 3- T细胞控制致病和保护反应的分子机制不同 在EAE开始和进展期间。我们将通过临床、细胞和分子分析来验证这一假设。 a）新的诱导型敲除小鼠，其中PD 1和/或LAG 3可以仅在Treg细胞中或仅在Treg细胞中暂时缺失。 CD 4 + Foxp 3- T细胞和B）给予PD 1和/或LAG 3阻断抗体的小鼠。我们将评估选定的目标 从我们的研究中定义了PD 1/LAG 3协调控制细胞命运和功能的机制。目标1： PD 1/LAG 3在Treg和自身反应性CD 4 + FoxP 3- T细胞中触发了哪些细胞和分子事件 在EAE发作期间删除或阻断？我们假设PD 1/LAG 3相互作用对Treg有不同的影响， vs. EAE起始期间的CD 4 + FoxP 3- T细胞。我们将定义PD 1和/或LAG 3缺失如何影响自身反应性 CD 4 + Foxp 3- T细胞活化/分化和Treg活化/功能，以及LAG 3/PD 1的综合作用 封锁目的2：在Treg和自身反应性CD 4 + Foxp 3效应子中触发了哪些细胞和分子事件 EAE发作后PD 1/LAG 3缺失或阻断Teff细胞？我们预测，PD 1/LAG 3相互作用可能不同， 在EAE的起始期与效应期，因为PD 1可以调节T细胞分化命运和效应子应答。 我们将定义PD 1和/或LAG 3缺失如何影响Teff和Treg细胞以及PD 1/LAG 3的综合效应。 封锁项目1将与项目2和项目3合作，比较PD 1/LAG 3中断对耐受性的影响 和T细胞耗竭：EAE中的CD 4 + FoxP 3- T细胞与癌症（项目2），以及EAE中的CD 4 + Teff与CD 8 + T细胞 癌症（项目2）和慢性感染（项目3）。Project 1将与Core A交互以交换数据，Core B用于获得独特的小鼠品系，核心C用于转录分析，核心D用于免疫病理学研究。

PROJECT SUMMARY PD1 and LAG3 play critical roles in regulating T cell tolerance and autoimmunity. Combined genetic deficiency of PD1 and LAG3 in mice results in lethal systemic autoimmunity, demonstrating PD1/LAG3 synergies in tolerance. The goal of Project 1 is to determine mechanisms by which PD1 and LAG3 control Treg and selfreactive CD4+ FoxP3– T cells in tolerance and autoimmunity. These are issues of fundamental and clinical significance, as PD1 and LAG3 are key mediators of T cell exhaustion and promising therapeutic targets, and there is a growing appreciation of immune-related adverse events in cancer patients treated with checkpoint blockade. A deeper understanding of how PD1 and LAG3 work together to mediate tolerance may enable optimal effective PD1 and LAG3 cancer therapies, while minimizing autoimmunity. In contrast to exacerbated EAE in Pdcd1–/– mice, our preliminary data show that mice lacking PD1 in Treg develop ameliorated EAE. Conversely, LAG3 deletion in Treg increased EAE severity, while combined PD1/LAG3 deletion in Treg reduced EAE. These findings highlight the need to determine how PD1/LAG3 signals are integrated in different cell types to understand how perturbing these pathways impacts protective Treg and pathogenic self-reactive CD4+ FoxP3– T cells. Based on these data, we hypothesize that synergy between PD1 and LAG3 operates by cellular and molecular mechanisms that differ in Treg and CD4+ FoxP3– T cells to control pathogenic and protective responses during EAE initiation and progression. We will test this hypothesis by clinical, cellular, and molecular analyses of a) novel inducible knockout mice where PD1 and/or LAG3 can be temporally deleted only in Treg cells or only in CD4+ Foxp3– T cells and b) mice given PD1 and/or LAG3 blocking antibodies. We will evaluate select targets from our studies to define mechanisms by which PD1/LAG3 coordinate to control cell fate and function. Aim 1: What cellular and molecular events are triggered in Treg and self-reactive CD4+ FoxP3– T cells by PD1/ LAG3 deletion or blockade during EAE onset? We hypothesize that PD1/LAG3 interactions have distinct effects on Treg vs. CD4+ FoxP3– T cells during EAE initiation. We will define how PD1 and/or LAG3 deletion affects self-reactive CD4+ Foxp3– T cell activation/differentiation and Treg activation/function, and integrated effects of LAG3/PD1 blockade. Aim 2: What cellular and molecular events are triggered in Treg and self-reactive CD4+ Foxp3– effector (Teff) cells by PD1/LAG3 deletion or blockade after EAE onset? We predict that PD1/LAG3 interactions may differ in initiation vs. effector phases of EAE, since PD1 can regulate T cell differentiation fates and effector responses. We will define how PD1 and/or LAG3 deletion impacts Teff and Treg cells and integrated effects of PD1/LAG3 blockade. Project 1 will collaborate with Projects 2 and 3 to compare effects of PD1/LAG3 disruption in tolerance and T cell exhaustion: CD4+ FoxP3– T cells in EAE vs. cancer (Project 2), and CD4+ Teff in EAE vs. CD8+ T cells in cancer (Project 2) and chronic infection (Project 3). Project 1 will interact with Core A to exchange data, Core B to obtain unique mouse strains, Core C for transcriptional analyses, and Core D for immunopathology studies.