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.

抽象的 免疫检查点 (ICP) 对免疫防御、监视、 通过配体-受体相互作用进行调节和自我耐受。 ICPs 存在于膜结合 和体内可溶形式。抑制性和刺激性膜结合 ICP (mICP) 之间的不平衡 外周血和组织中的恶性细胞和免疫细胞已被充分记录。封锁 CTLA-4、PD-1 和 PD-L1 等抑制性 mICP 已成为晚期晚期患者的革命性治疗方法 黑色素瘤等恶性肿瘤的各个阶段。然而，两者的起源、调控和生物学意义 mICPs 和可溶性 ICPs (sICPs) 在中枢神经系统 (CNS) 中的作用尚未得到系统的研究 研究过。我们使用多重免疫分析同时量化 16 种 sICP 的浓度 来自 33 名 HIV 阴性者 (HNI) 和 105 名 HIV 感染者的脑脊液 (CSF) 样本 (PLWH)，包括 80 例 HIV 相关神经认知障碍 (HAND) 病例和 25 例无 HAND 病例。 我们发现 10 个 sICP（sHVEM、sCD27、sGITR、sICOS、sLAG-3、sPD-1、sTIM-3、sBTLA、sCD28 和 sCD40） 在所有脑脊液样本中均检测到，其中大多数在感染者中高度升高。引人注目的是， 与没有 HAND 的 PLWH 相比，HAND 中的 CSF sHVEM 显着增加。这 10 个 sICP 在恒河猴 (RM) 的脑脊液样本中也可检测到。免疫组织化学 (IHC) 分析 来自携带或不携带猿猴免疫缺陷病毒的 RM 的脑组织（额叶皮层和海马） (SIV) 感染表明 mHVEM 在神经元上以基础水平表达，但在星形胶质细胞或星形胶质细胞上不表达。 在 SIV 感染的 RM (SIV/RM) 中，小胶质细胞和 mHVEM 表达显着增加。此外，我们 最近报道，U87MG 细胞（人类星形胶质细胞系）中的 PD-L1 表达可以稳健地 由 IFN-γ/IL-1β/TNF-α 混合物诱导，诱导的 PD-L1 控制 MCP-1 的产生（最 通过 PD-L1 内在信号传导调节单核细胞/巨噬细胞迁移的重要趋化因子。因此， 我们假设 sICPs 和 mICPs 在 PLWH 的 CNS 中失调，这与 手神经发病机制。我们有两个具体目标来检验这个中心假设：(1) 确定 sICP PLWH 和 SIV/RM 中枢神经系统的概况和生物学功能，以及 (2) 表征细胞生态位 ICP受体/配体阳性神经细胞和浸润性免疫细胞在大脑中的空间关联 PLWH 和 SIV/RM。我们的结果将深入了解 ICP 在 CNS 神经免疫中的作用 有或没有 HAND 的 PLHIV 的失调和神经炎症。 sICP 的 CSF 水平和概况 例如 sHVEM 可以作为 HAND 严重程度和进展的生物标志物。