Investigating the Repertoires and Functions of T Cells that Help Autoreactive B Cells in Lupus

狼疮是一种主要影响年轻女性的毁灭性疾病。狼疮自身免疫是 许多不同的自身免疫性疾病的原型，共同影响3-5%的人口， 是的。T细胞的激活依赖于B细胞和先天免疫感应，是关键 促进疾病。尽管T细胞在驱动几乎所有方面的系统性免疫方面都是至关重要的， 自身免疫性疾病，它们的身份，特异性，功能和命运仍然非常不明确。Au- 抗反应性T细胞为自身反应性B细胞提供帮助，并通过靶向T细胞引起组织损伤。 器官浸润虽然我们对生物多样性的特殊性、调节和命运有着深刻的理解， 对于自身反应性B细胞来说，同样的情况不能说是自身反应性T细胞。我们尤其 我想阐明自身反应性T细胞如何受到自身耐受机制的调节，以及如何， 一旦激活，它们就对自身免疫的各个方面起作用。 通过目前的提案，我们希望在这一重要而又不足的领域取得重大进展- 简易爆炸装置区。我们将首先使用一种新的策略来分离自体反应性T细胞的新克隆， 识别来自正常和自身免疫性易感遗传基因的核组分（“ANA T细胞”） 背景（目的1A）;这将提供对自身免疫B辅助T细胞库的见解 细胞库然后，我们将选择的T细胞的TCR克隆到逆转录病毒表达载体中， 制造“逆转录”（Rg）小鼠。然后我们将确定（目标1B）这些自身反应细胞如何 发展，受到各种耐受机制的影响，并且可能是自发的 激活这将根据T细胞的起源以及背景进行研究 其中T细胞表达（正常或自身免疫背景）。在目标1（C）中，我们将 将选定的TCR转化为完全成熟的TCR转基因（Tg）小鼠，这将是 更强大和详细的机制研究。在目标2中，我们将使用这些Tg小鼠T细胞， 研究活化、T-B相互作用和Toll样受体（TLR）依赖性的机制。这 将由一系列混合骨髓嵌合体完成，这将使我们能够询问命运， ANA T细胞在更自然的环境中发育和运作 在由嵌合体中的其他细胞驱动的多克隆自身免疫的情况下， 同源BCR的表达以及TLR信号传导能力。 在这些研究结束时，我们不仅将拥有一套真正独特的新工具， 正常和自身免疫背景下的克隆形式、Rg小鼠和Tg小鼠，但我们将有 在了解ANA T细胞如何激活方面非常显着地推进了该领域 它们如何促进疾病，以及它们如何与先天免疫信号相互作用。

Lupus is a devastating disease predominantly affecting young women. Lupus autoimmunity is prototypical of many different autoimmune diseases, which together affects 3-5% of the popula- tion. T cells, whose activation depends on both B cells and innate immune sensing, are pivotal in promoting disease. Despite that T cells are critical in driving virtually all aspects of systemic autoimmune disease, their identity, specificity, function and fate remain very poorly defined. Au- toreactive T cells provide help to autoreactive B cells and also cause tissue damage by target organ infiltration. While we have a deep understanding of the specificity, regulation and fate of autoreactive B cells, the same cannot be said for autoreactive T cells. In particular, we would like to elucidate how self-reactive T cells are regulated by self-tolerance mechanisms and how, upon activation, they contribute to various aspects of autoimmunity. With the current proposal we hope to make significant inroads into this important yet understud- ied area. We will first use a novel strategy to isolate new clones of autoreactive T cells that rec- ognize nuclear components (“ANA T cells”) from both normal and autoimmune prone genetic backgrounds (Aim 1A); this will provide insights into the repertoire of autoimmune B-helper T cell repertoire. We will then clone the TCRs of selected T cells into retroviral expression vectors to make “retrogenic” (Rg) mice. We will then determine (Aim 1B) how these autoreactive cells develop, are subject to a variety of tolerance mechanisms, and are potentially spontaneously activated. This will be studied according to both the origin of the T cell as well as the context in which the T cell is expressed (normal or autoimmune background). Finally in Aim 1(C), we will convert selected TCRs into full-fledged TCR transgenic (Tg) mice, which will be required for more robust and detailed mechanistic studies. In Aim 2 we will then use these Tg mice T cells to study mechanisms of activation, T-B interaction and Toll-like receptor (TLR) dependence. This will be done by a series of mixed bone marrow chimeras that will allow us to interrogate the fate and function of ANA T cells in a more natural milieu in which the T cells will develop and operate in the context of polyclonal autoimmunity driven by the other cells in the chimera that will vary in expression of cognate BCRs as well as TLR signaling capability. At the conclusion of these studies we will not only have a truly unique set of new tools in the form of clones, Rg mice and Tg mice on normal and autoimmune backgrounds, but we will have very significantly advanced the field in terms of understanding how ANA T cells are activated and regulated, how they promote disease, and how they interact with innate immune signals.