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

研究帮助狼疮中自身反应性 B 细胞的 T 细胞的组成和功能

• 批准号: 10058242
• 负责人: MARK J SHLOMCHIK
• 金额: $ 38.28万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-22 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10058242
• 关键词: Address; Affect; Antigen-Presenting Cells; Antinuclear Antibodies; Area; Attention; Autoantibodies; Autoimmune; Autoimmune Diseases; Autoimmunity; Automobile Driving; B-Lymphocytes; Biological Assay; Bone Marrow; Bypass; Cell Communication; Cells; Chimera organism; Clone Cells; DNA; Data; Dependence; Disease; Flow Cytometry; Frequencies; Funding; Genetic; Goals; Helper-Inducer T-Lymphocyte; Immune; Immune signaling; Immunoglobulin G; Inbred BALB C Mice; Infiltration; Kidney; Lupus; Methods; Molecular; Mouse Strains; Mus; Nuclear; Nucleic Acids; Organ; Pathogenesis; Pathogenicity; Peripheral; Play; Population; Production; RNA; Reagent; Receptor Signaling; Regulation; Regulatory T-Lymphocyte; Reporting; Role; Self Tolerance; Series; Specificity; T-Cell Development; T-Cell Receptor; T-Lymphocyte; TLR7 gene; Testing; Tissues; Toll-like receptors; Transgenic Mice; Transgenic Organisms; Tumor-infiltrating immune cells; United States National Institutes of Health; Work; anergy; anti-IgG; autoreactive B cell; autoreactive T cell; autoreactivity; central tolerance; expression vector; in vivo; innate immune sensing; insight; interest; new therapeutic target; novel; novel strategies; novel therapeutics; receptor expression; sensor; systemic autoimmune disease; tool; virtual; young woman

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.

狼疮是一种毁灭性的疾病，主要影响年轻女性。狼疮自身免疫性是 许多不同的自身免疫性疾病的典型，这些疾病加起来影响了3%-5%的人口- 提顿。T细胞的激活依赖于B细胞和先天免疫感知，它是关键 在促进疾病方面。尽管如此，T细胞在推动全身性疾病的几乎所有方面都是至关重要的 对于自身免疫性疾病，它们的身份、特异性、功能和命运仍然很不清楚。Au- TO反应性T细胞为自身反应性B细胞提供帮助，也可通过靶点造成组织损伤 器官渗入。虽然我们对这一现象的特殊性、规律性和命运有着深刻的理解 自体反应性B细胞，但自体反应性T细胞就不是这样了。尤其是，我们会 我想阐明自我反应性T细胞是如何受到自我耐受机制的调节的， 一旦激活，它们对自身免疫的各个方面都有贡献。 通过目前的建议，我们希望在这一重要但被理解的领域取得重大进展- 简易爆炸装置区域。我们将首先使用一种新的策略来分离新的自身反应性T细胞克隆，这些克隆可以重新识别- 识别正常和自身免疫易感基因的核成分(“ANA T细胞”) 背景(目标1A)；这将提供对自身免疫性B辅助T细胞的曲目的见解 细胞谱系。然后，我们将把选定T细胞的TCR克隆到逆转录病毒表达载体中 制造“逆转录基因”(Rg)小鼠。然后我们将确定(目标1B)这些自体反应细胞是如何 发展，受制于各种耐受机制，并可能自发地 激活了。这将根据T细胞的起源以及 T细胞的表达(正常或自身免疫背景)。最后，在目标1(C)中，我们将 将选定的TCR转化为成熟的TCR转基因(TG)小鼠，这将是 更有力、更详细的机械论研究。在目标2中，我们将使用这些转基因小鼠的T细胞 研究激活、T-B相互作用和Toll样受体(TLR)依赖机制。这 将由一系列混合的骨髓嵌合体完成，这将使我们能够询问命运 ANA T细胞在更自然的环境中的功能，在这种环境中，T细胞将发展和运作 在由嵌合体中的其他细胞驱动的多克隆自身免疫的背景下， 同源BCR的表达以及TLR信号转导能力。 在这些研究的结论中，我们不仅将拥有一套真正独特的新工具 克隆的形式，Rg小鼠和Tg小鼠在正常和自身免疫背景下，但我们将有 在了解ANA T细胞如何激活方面，该领域取得了非常显著的进步 以及它们如何促进疾病，以及它们如何与先天免疫信号相互作用。