Targeting follicular helper CD4 T cells in SLE

项目摘要/摘要 这项提案旨在描述TFH细胞的特征，TFH细胞是一种对疾病发展和 维持在SLE中，并检验SLE-Tfh对代谢抑制唯一敏感的新假设，a 可用于治疗干预的特征。这项提议的科学前提是基于 三条证据：1.红斑狼疮中TFH细胞在产生致病自身抗体中的必要作用 已经在SLE患者和小鼠模型中得到了很好的证实；2.免疫诱导的TFH细胞 代谢处于静止状态，这一事实已经得到证实，表明它们不受葡萄糖抑制剂的影响 我们有强有力的初步数据表明，来自四个人的SLE-Tfh细胞 不同的狼疮小鼠模型对葡萄糖敏感。 我们提出的假设是，支持自身抗体产生的自发Tfh细胞(SLE-Tfh)具有 与Tfh细胞不同的代谢，提供对病原体的保护性体液免疫(TD-Tfh) 狼疮或正常小鼠。基于狼疮易感小鼠和狼疮易感小鼠的CD4+T细胞功能和代谢的相似性 我们还假设，在SLE患者中，扩增的Tfh细胞也有不同的 新陈代谢水平高于健康对照组(HCS)。因此，我们预测靶向TFH细胞代谢 提供了一种有效的方法来治疗狼疮而不影响患者对TD的保护- 病原体。我们建议使用细胞和分子免疫学方法来检验这些假说 小鼠模型和人类外周血淋巴细胞模型，具有以下三个特定目的 1.确定SLE-Tfh细胞与TD-Tfh细胞的分子和代谢特征 由依赖TD的AGS(PR8流感病毒和NP-OVA)在狼疮小鼠和B6对照组中诱发。这个 自发性Tfh细胞的免疫表型、解剖定位、基因表达和代谢谱 将TC和B6.lpr小鼠与流感特异性I-A(B)NP-四聚体阳性Tfh细胞在TC，B6.lpr中进行比较 感染PR8病毒的B6小鼠。除了这些多克隆T细胞模型外，我们还将使用一种 携带SLE易感等位基因的OVA特异性OT-II T细胞的转移模型 TFH细胞(9)扩增成NP-OVA免疫小鼠(10)。这一目标将定义功能差异 在两种不同TD免疫的狼疮模型中，TD-Tfh和SLE-Tfh细胞之间的差异。 2.研究小鼠SLE-Tfh和TD-Tfh细胞对葡萄糖抑制的反应。使用 与SA1相同的实验系统，我们将比较SLE-Tfh细胞和TD-Tfh细胞对 用2DG处理的小鼠的葡萄糖抑制，2DG是一种葡萄糖类似物，可以阻断糖酵解的第一反应。 3.比较SLE和SLE患者外周血中cTfh细胞的分子和代谢特征。 HCS，以及它们对二甲双胍的反应。我们推测系统性红斑狼疮中cTfh细胞的增殖 患者在很大程度上是由自体抗原驱动的，并且SLE-cTfh细胞与小鼠SLE-Tfh细胞具有共同的功能特征 细胞。另一方面，来自HCS的cTfh细胞已经被TD-AGS大量诱导，HC-cTfh细胞应该 与小鼠TD-Tfh细胞重叠。我们将比较两者的免疫表型和基因表达。 TFH细胞的类型，以及它们对代谢抑制剂二甲双胍的体外反应。 我们的第一个目标是促进我们对SLE-Tfh细胞的理解，并最终推进对SLE-Tfh细胞的治疗 狼疮，基于离散的、可实现的目标，专注于一个细胞群体。这个项目将推进我们的 对疾病机制的理解，并产生具有很高翻译潜力的结果。

Project Summary/Abstract This proposal intends to characterize Tfh cells, a population that is critical for disease development and maintenance in SLE and test the novel hypothesis that SLE-Tfh are uniquely sensitive to metabolic inhibition, a feature that can be exploited to therapeutic intervention. The scientific premise of this proposal is based on three lines of evidence: 1. The necessary role of Tfh cells for the production of pathogenic autoAbs in lupus has been well-established in SLE patients and mouse models; 2. Tfh cells induced by immunization are metabolically quiescent, a fact that we have confirmed showing that they are not affected by glucose inhibitors in lupus-prone and control mice; and 3. We have strong preliminary data showing that SLE-Tfh cells from four different mouse models of lupus are sensitive to glucose. We propose the hypothesis that spontaneous Tfh cells supporting the production of autoAbs (SLE-Tfh) have a different metabolism than Tfh cells providing protective humoral immunity against pathogens (TD-Tfh) in either lupus or normal mice. Based on similarities of CD4+ T cell functions and metabolism in lupus-prone mice and SLE patients, we also hypothesize that the expanded Tfh cells in SLE patients also have a different metabolism than that of health controls (HCs). Consequently, we predict that targeting Tfh cellular metabolism provides an effective approach to treat lupus without compromising the patients’ protection against TD- pathogens. We propose to test these hypotheses using cellular and molecular immunology approaches in mouse models as well as with human PBLs with the three following specific aims 1. To define the molecular and metabolic signatures of SLE-Tfh as compared to TD-Tfh cells elicited by TD-dependent Ags (PR8 flu virus and NP-OVA) in lupus mice and B6 controls. The immunophenotypes, anatomical location, gene expression, and metabolic profile of spontaneous Tfh cells in TC and B6.lpr mice will be compared to that of flu-specific I-A(b) NP-tetramer positive Tfh cells in TC, B6.lpr and B6 mice infected with PR8 virus. In addition to these polyclonal T cell models, we will use an adoptive transfer model of OVA-specific OT-II T cells carrying the Sle1 lupus susceptibility allele that favors the expansion of Tfh cells (9) into NP-OVA immunized mice (10). This aim will define the functional differences between TD-Tfh and SLE-Tfh cells in two models of lupus and with two different TD-immunizations. 2. To define the response of SLE-Tfh and TD-Tfh cells to glucose inhibition in the mouse. Using the same experimental systems as in SA1, we will compare the responses of SLE-Tfh cells and TD-Tfh cells to glucose inhibition in mice treated with 2DG, a glucose analog that blocks the first reaction of glycolysis. 3. To compare the molecular and metabolic signatures of circulating cTfh cells in SLE patients and HCs, as well as their response to metformin. We hypothesize that the expansion of cTfh cells in SLE patients is largely driven by autoAgs, and that SLE-cTfh cells share functional signatures with murine SLE-Tfh cells. On the other hand, cTfh cells from HCs have been largely induced by TD-Ags, and HC-cTfh cells should overlap with murine TD-Tfh cells. We will compare the immunophenotypes and gene expression of these two types of Tfh cells, as well as their response to the metabolic inhibitor metformin in vitro. Our first goal is to advance our understanding of SLE-Tfh cells, and ultimately to advance the treatment of lupus, based on discrete, achievable goals focusing on one cell population. This project will advance our understanding of disease mechanisms, and yield results with a high translational potential.