Immune checkpoints in the CNS and HIV-associated neurocognitive disorder

中枢神经系统和艾滋病毒相关神经认知障碍中的免疫检查点

• 批准号: 10889463
• 负责人: Yunlong Liu
• 金额: $ 82.37万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10889463
• 关键词: Address; Affect; Alternative Splicing; Astrocytes; Bar Codes; Binding; Biological; Biological Markers; Biological Process; Biological Response Modifiers; Brain; CD8-Positive T-Lymphocytes; CRISPR/Cas technology; CTLA4 gene; Cell Line; Cells; Central Nervous System; Cerebrospinal Fluid; Circulation; Clinical; DNA; Data; Detection; Enzyme-Linked Immunosorbent Assay; Exhibits; Frequencies; Gene Expression Profile; Genes; Growth Factor; HIV; HIV Seronegativity; HIV-associated neurocognitive disorder; Hippocampus; Homeostasis; Human; Image; Immune; Immunity; Immunoassay; Immunohistochemistry; Immunoprecipitation; Individual; Inflammatory; Interferon Type II; Interleukin-1 beta; Lead; Ligands; Macaca mulatta; Macrophage; Malignant Neoplasms; Measures; Membrane; Messenger RNA; Microglia; Neuroimmune system; Neurologic Symptoms; Neurons; Neuropathogenesis; Pathogenesis; Pathway interactions; Peripheral; Persons; Production; Protein Isoforms; Protein Secretion; Proteins; RNA Splicing; Regulation; Reporting; Repression; Research; Resolution; Reverse Transcriptase Polymerase Chain Reaction; Role; SIV; Sampling; Self Tolerance; Severities; Signal Transduction; T cell regulation; TNF gene; Tail; Techniques; Testing; Time; Tissue Sample; Tissues; Variant; Virus Diseases; antibody conjugate; brain cell; brain tissue; cancer cell; cell type; checkpoint receptors; chemokine; clinically relevant; cytokine; digital imaging; frontal lobe; immune cell infiltrate; immune checkpoint; in vivo; indexing; innovation; insight; melanoma; migration; monocyte; multiplex assay; neurocognitive disorder; neuroinflammation; peripheral blood; programmed cell death ligand 1; programmed cell death protein 1; receptor; single nucleus RNA-sequencing; spatial relationship; tumor

Abstract Immune checkpoints (ICPs) exert inhibitory or stimulatory effects on immune defense, surveillance, regulation, and self-tolerance through their ligand-receptor interactions. ICPs exist in both membrane-bound and soluble forms in vivo. Imbalances between inhibitory and stimulatory membrane-bound ICPs (mICPs) in malignant cells and immune cells in the peripheral blood and tissues have been well documented. Blockade of inhibitory mICPs such as CTLA-4, PD-1, and PD-L1 has become a revolutionary treatment for advanced stages of malignancies such as melanoma. However, the origin, regulation, and biological significance of both mICPs and soluble ICPs (sICPs) in the central nervous system (CNS) have not yet been systematically studied. We used a multiplex immunoassay to simultaneously quantify the concentrations of 16 sICPs in cerebrospinal fluid (CSF) samples from 33 HIV-negative individuals (HNIs) and 105 people living with HIV (PLWH), including 80 cases with HIV-associated neurocognitive disorder (HAND) and 25 cases without HAND. We found that 10 sICPs (sHVEM, sCD27, sGITR, sICOS, sLAG-3, sPD-1, sTIM-3, sBTLA, sCD28, & sCD40) were consistently detected in all CSF samples, and most of them were highly elevated in PLWH. Strikingly, CSF sHVEM was significantly increased in HAND when compared to PLWH without HAND. These 10 sICPs were also detectable in the CSF samples from rhesus macaques (RMs). Immunohistochemistry (IHC) analysis of brain tissues (frontal cortex and hippocampus) from RMs with and without simian immunodeficiency virus (SIV) infection showed that mHVEM was expressed at a basal level on neurons, but not on astrocytes or microglia, and mHVEM expression was highly increased in SIV-infected RMs (SIV/RMs). Furthermore, we have recently reported that PD-L1 expression in U87MG cells (a human astrocyte cell line) can be robustly induced by a mixture of IFN-γ/IL-1β/TNF-α, and induced PD-L1 controls production of MCP-1 (the most important chemokine that regulates migration of monocytes/macrophages) via PD-L1 intrinsic signaling. Thus, we hypothesize that both sICPs and mICPs are dysregulated in the CNS of PLWH, which are associated with HAND neuropathogenesis. We have two Specific Aims to test this central hypothesis: (1) To determine sICP profiles and biological functions in the CNS of PLWH and SIV/RMs, and (2) To characterize the cellular niches and spatial associations of ICP receptor/ligand-positive neural cells and infiltrating immune cells in the brain of PLWH and SIV/RMs. Our results will provide insights into the roles of ICPs in the CNS neuroimmune dysregulation and neuroinflammation of PLHIV with and without HAND. The CSF levels and profiles of sICPs such as sHVEM can potentially be used as a biomarker for HAND severity and progression.

摘要 免疫检查点(ICP)在免疫防御、监视、 通过配体-受体的相互作用调节和自我耐受。ICP存在于两种膜结合体中 和体内可溶的形式。抑制性和刺激性膜结合蛋白(MICPs)之间的失衡 外周血和组织中的恶性细胞和免疫细胞已经被很好地记录下来。封锁 抑制性mICPs如CTLA-4、PD-1和PD-L1已成为晚期癌症的革命性治疗方法 恶性疾病的分期，如黑色素瘤。然而，两者的起源、调控和生物学意义 中枢神经系统(CNS)中的MICPs和可溶性ICPs(SICPs)尚未得到系统的研究 学习。我们使用多重免疫测定法同时定量测定了16种SICP的浓度 33例HIV阴性者和105例HIV感染者的脑脊液样本 (PLWH)，包括80例HIV相关神经认知障碍(手)和25例无手。 我们发现了10个SICP(sHVEM、sCD27、sGITR、SiCOS、熔渣-3、SPD-1、STIM-3、sBTLA、sCD28和sCD40)。 在所有脑脊液标本中持续检测到，其中大部分在PLWH中高度升高。令人惊讶的是， 与非手部PLWH相比，手部CSFsHVEM显著增加。这10个SICP 在恒河猴(RMS)的脑脊液样本中也能检测到。免疫组织化学(IHC)分析 大脑组织(额叶皮质和海马体)中携带和不携带猴免疫缺陷病毒的RMS (SIV)感染显示mHVEM在神经元上有基础水平的表达，而在星形胶质细胞或 在SIV感染的RMS(SIV/RMS)中，mHVEM的表达显著增加。此外，我们 最近有报道称，PD-L1在U87 MG细胞(一种人星形胶质细胞系)中的表达可以很强 由干扰素-γ/白介素1-β/肿瘤坏死因子-α的混合物诱导，以及诱导的PD-L1控制单核细胞趋化蛋白-1的产生(最 通过PD-L1内在信号调节单核/巨噬细胞迁移的重要趋化因子)。因此， 我们假设SICP和MICP在PLWH的中枢神经系统中调节失调，这与 手部神经发病机制。我们有两个具体的目的来检验这个中心假设：(1)确定SiCp PLWH和SIV/RMS中枢神经系统的分布和生物学功能，以及(2)细胞生态位的特征 脑内ICP受体/配体阳性神经细胞与浸润性免疫细胞的空间相关性 PLWH和SIV/RMS。我们的结果将为深入了解ICPs在中枢神经免疫中的作用提供依据 有手和无手的PLHIV的调节失调和神经炎症。SICP患者脑脊液的水平和分布 例如sHVEM有可能被用作手部严重程度和进展的生物标记物。