B cells alter adaptive autoimmunity to protect from ischemic injury after stroke

B 细胞改变适应性自身免疫以预防中风后缺血性损伤

• 批准号: 9385820
• 负责人: Ann Marie Stowe
• 金额: $ 9.29万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-01 至 2018-08-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9385820
• 关键词: 3-Dimensional; Acute; Adoptive Transfer; Antigens; Attenuated; Autoimmune Responses; Autoimmunity; Axon; B-Lymphocytes; Behavioral; Behavioral Assay; Blood - brain barrier anatomy; Brain; Brain Injuries; Brain imaging; Cause of Death; Cell Communication; Cells; Clinical Treatment; Clinical Trials; Coculture Techniques; Data; FDA approved; GTP-Binding Protein alpha Subunits, Gs; Glucose; Goals; Grant; Harvest; Hypoxia; Immune; Immune response; In Vitro; Infarction; Inflammation; Inflammatory; Inflammatory Response; Injury; Interferon Type II; Interferon-alpha; Interleukin-10; Intervention; Ischemia; Knockout Mice; Leukocytes; Longitudinal Studies; Magnetic Resonance Imaging; Measures; Mediating; Mediator of activation protein; Methodology; Methods; Modeling; Morbidity - disease rate; Motor; Mus; Myelin; Neuraxis; Neuronal Injury; Neuronal Plasticity; Neurons; Oxygen; Pathology; Patients; Pharmaceutical Preparations; Phenocopy; Phenotype; Population; Pre-Clinical Model; Production; Recombinant interferon beta-1b; Recovery; Recovery of Function; Recruitment Activity; Regulatory T-Lymphocyte; Research; Research Personnel; Role; Stroke; T-Lymphocyte; Testing; Th1 Cells; Therapeutic; Three-Dimensional Imaging; Time; Treatment Efficacy; Wild Type Mouse; Work; adaptive immune response; adaptive immunity; autoreactive T cell; autoreactivity; base; betacell therapy; cohort; copolymer 1; cytokine; deprivation; efficacy testing; excitotoxicity; immunoregulation; immunosuppressed; improved; in vitro testing; injured; interleukin-10 receptor; macrophage; male; migration; monocyte; mortality; motor deficit; multiple sclerosis patient; neglect; neurogenesis; neuroprotection; neurovascular; neurovascular injury; neutrophil; novel; post stroke; preclinical study; preconditioning; prevent; public health relevance; repaired; response; secondary infection; stroke recovery; tomography; two-photon

 DESCRIPTION (provided by applicant): More than 800,000 people in the U.S. will suffer from stroke each year, yet there remains only one FDA- approved acute intervention. Clinical trials to minimize post-stroke inflammation failed, highlighting the need to better understand inflammatory and neuroprotective mechanisms within the central nervous system (CNS). Repetitive hypoxic preconditioning (RHP) naturally protects from stroke for months beyond treatment, minimizing infarct volumes, blood-brain barrier (BBB) disruption, and monocyte, T cell, and neutrophil diapedesis into the ischemic brain. In contrast, RHP specifically induces an immunosuppressed B cell phenotype that is enhanced in the injured CNS of preconditioned mice following stroke. The adoptive transfer of RHP-treated B cells (B(RHP)cells) 6h after transient stroke in male mice reduced infarct volumes and improved functional recovery compared to PBS- or wild type (WT) B cell-treated controls. B(RHP)cell therapy also suppressed a post-stroke autoimmune response to CNS-derived neuronal and myelin antigen (Ag), a novel finding for adaptive autoimmunity that has only been studied in preclinical models of stroke in the context of secondary infection. The goal of this New Investigator grant is to determine if B(RHP)cells, as part of a transferrable adaptive immunity to mild hypoxia, mediate endogenous protection through the production of interleukin 10 (IL10), a cytokine that promotes neuronal repair, as well as interferon (IFN)-γ, a cytokine that converts effector (i.e. pro-inflammatory, Th1) T cells to regulatory T cells (Tregs) that suppress other neuropathological immune cells. Specific Aim 1 will test the hypothesis that B cells confer acute neuroprotection through the production of IL10 and direct neuronal interaction. We will confirm, using state-of-the-art 3-D serial two-photon tomography for whole brain imaging, MRI, and behavioral assays, that IL10- enhanced B(RHP)cells colocalize to a higher number of surviving neurons in the ischemic hemisphere to improve long-term functional recovery. We will also confirm the role of B(RHP)cell-derived IL10 on neuronal viability and axon outgrowth in vitro following oxygen-glucose deprivation (OGD), with B(RHP)cells harvested from IL10-null and WT mice. Specific Aim 2 will test the hypothesis that B(RHP)cells from WT but not IFN-γ-null mice will induce a novel Treg population to suppress pro-inflammatory autoreactive responses from Th1 cells. The transfer of Tregs exposed in vitro to B(RHP)cells will also promote long-term neurovascular and functional recovery (Aim 1A methods), in addition to suppressing CNS-derived autoreactivity. Finally, Glatiramer Acetate, or Copaxone(r), is an FDA-approved drug that upregulates B cell-derived IL10 and Treg conversion in patients with multiple sclerosis. Specific Aim 3 will test the hypothesis that WT B cells treated with Copaxone will upregulate IL10 and IFN-γ to phenocopy B(RHP)cell-mediated protection. We will test the efficacy of Copaxone-treated B cells on neuronal viability in vitro following OGD (Aim 1B methods). Copaxone- or control Betaseron-treated B cells will be adoptively transferred post-stroke to determine efficacy on long-term recovery (Aim 1A methods).

 描述（由申请人提供）：美国每年将有超过80万人患有中风，但目前只有一种FDA批准的急性干预措施。减少中风后炎症的临床试验失败了，突出了更好地了解中枢神经系统（CNS）内炎症和神经保护机制的必要性。重复性低氧预处理（RHP）在治疗后的数月内自然地防止中风，最大限度地减少梗死体积、血脑屏障（BBB）破坏以及单核细胞、T细胞和中性粒细胞渗出到缺血性脑中。相反，RHP特异性诱导免疫抑制的B细胞表型，其在中风后预适应小鼠的受损CNS中增强。与PBS或野生型（WT）B细胞处理的对照组相比，在雄性小鼠短暂性卒中后6小时过继转移RHP处理的B细胞（B（RHP）细胞）减少了梗死体积并改善了功能恢复。B（RHP）细胞疗法还抑制了对CNS衍生的神经元和髓鞘抗原（Ag）的中风后自身免疫应答，这是适应性自身免疫的一个新发现，仅在继发感染的中风临床前模型中进行了研究。这项新研究者基金的目标是确定B（RHP）细胞作为对轻度缺氧的可转移适应性免疫的一部分，是否通过产生白细胞介素10（IL 10）（一种促进神经元修复的细胞因子）以及干扰素（IFN）-γ（一种将效应细胞转化为细胞因子）来介导内源性保护。（即促炎性，Th 1）T细胞转化为抑制其他神经病理免疫细胞的调节性T细胞（TcR）。具体目标1将检验B细胞通过产生IL 10和直接神经元相互作用赋予急性神经保护的假设。我们将使用最先进的3-D连续双光子断层扫描进行全脑成像、MRI和行为分析，证实IL 10增强的B（RHP）细胞共定位于缺血半球中更多数量的存活神经元，以改善长期功能恢复。我们还将证实B（RHP）细胞衍生的IL 10在体外氧-葡萄糖剥夺（OGD）后对神经元活力和轴突生长的作用，其中B（RHP）细胞收获自IL 10缺失和WT小鼠。具体目标2将检验以下假设：来自WT而非IFN-γ-无效小鼠的B（RHP）细胞将诱导新的Treg群体，以抑制来自Th 1细胞的促炎性自身反应性应答。除了抑制CNS衍生的自身反应性外，将体外暴露的TdR转移至B（RHP）细胞还将促进长期神经血管和功能恢复（目的1A方法）。最后，Glatiramer Acetate或Copaxone（r）是FDA批准的药物，可上调多发性硬化症患者的B细胞来源的IL 10和Treg转化。具体目标3将检验用克帕松处理的WT B细胞将上调IL 10和IFN-γ对表型复制B（RHP）细胞介导的保护的假设。我们将在OGD后体外测试经克帕松处理的B细胞对神经元活力的功效（Aim 1 B方法）。将在中风后过继转移经克帕松或对照Betaseron处理的B细胞，以确定对长期恢复的功效（目的1A方法）。