CALPAIN ACTIVATION OF T CELLS IN DEMYELINATING DISEASE

钙蛋白酶激活 T 细胞在脱髓鞘疾病中的作用

• 批准号: 7590468
• 负责人: NAREN L BANIK
• 金额: $ 31.9万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-01-15 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7590468
• 关键词: Apoptotic; Astrocytes; Attenuated; Autoimmune Process; Axon; Biological Assay; CD4 Positive T Lymphocytes; Calcium; Calpain; Cell Death; Cell Nucleus; Cell Survival; Cell physiology; Cells; Cessation of life; DNA Fragmentation; Data; Demyelinating Diseases; Demyelinations; Digestion; Disease remission; Enzyme-Linked Immunosorbent Assay; Epitopes; Etiology; Event; Fluorescence; Fura-2; Generations; Human; IL8RA gene; IL8RB gene; Immigration; In Vitro; Individual; Inflammatory; Inflammatory Response; Interferon Type II; Interferons; Interleukin-10; Interleukin-12; Interleukin-13; Interleukin-2; Interleukin-4; Interleukin-5; Interleukin-8; Mediating; Membrane Potentials; Methods; Microglia; Multiple Sclerosis; Myelin; Myelin Basic Proteins; Nerve Degeneration; Neuraxis; Neuroglia; Neurons; Nuclear; Nuclear Translocation; Oligodendroglia; Pathogenesis; Patients; Peptide Hydrolases; Peptides; Peripheral Blood Mononuclear Cell; Plasma; Play; Polymerase Chain Reaction; Process; Production; Rattus; Relapse; Research Personnel; Role; SJA6017; Signal Pathway; Signaling Protein; T cell activating factor; T-Cell Activation; T-Lymphocyte; T-Lymphocyte Epitopes; TCF Transcription Factor; Techniques; Testing; Therapeutic; Tumor Necrosis Factor-alpha; Western Blotting; autoreactive T cell; calpain inhibitor; calpeptin; caspase-3; chemokine; chemokine receptor; cytochrome c; cytokine; functional restoration; immunogenic; migration; neuron loss; programs; release factor; restoration; treatment strategy

Strong evidence suggests that multiple sclerosis (MS) is mediated through an autoimmune attack by myelin-specific T cells causing damage to myelinated axons and cells. Understanding the underlying mechanisms by which T cells become activated, migrate into CMS,and exert damage is crucial to treating the demyelinating disease. Calpain, a calcium (Ca )-activated neutral protease, has been implicated in MS. This competing renewal application will determine whether alterations in calpain activity in relapse and remission MS patients are involved in T cell activation and migration, Th1/Th2 dysregulation, epitope spreading, and/or axonal damage and neuronal and glial cell death and, thus, examine the effects of calpain inhibition on these events. Preliminary data suggest that calpain expression and activity, concomitant with increased interieukin (IL)-2 production (Th1), are upregulated in MS plasma and peripheral blood mononuclear cells (PBMCs) supernatant of MS patients, compared to controls. The increase in IL-2 production in relapse and remission groups correlated with increased translocation of the nuclear factor NFATd. In contrast, IL-4 and IL-10 (Th2) levels were highest in MS remission compared to relapse. Calpain released from MS patient cells degraded MBP and calpain's activity was increased in primary astrocytes and microglia treated with interferon (IFN)-y (increased in MS). We hypothesize that increased calpain expression and activity in MS PBMCs and MBP-specific T cells in relapse and remission patients will correlate with changes in Th1/Th2 cytokine profiles, NFAT translocation, and IL-2 production; and that released calpain and other inflammatory factors from activated T cells will damage axons and neurons and degrade MBP into antigenic peptides. Calpain inhibitor treatment will inhibit T cells activation and migration, reduce pro-inflammatory response, and attenuate cell death and axon damage. These hypotheses will be tested by the following specific aims: (1) examine the correlation between calpain expression and activity and levels of Th1/Th2 cytokines in MS PBMCs and MBP-specific T cells, and plasma, compared to controls; (2) determine the mechanisms of activation and migration of MS PBMCs and MBP-specific T cells and examine the effects of calpain inhibition; determine whether MBP digestion into immunogenic peptides is calpain-dependent and if the peptides sufficiently activate MS T cells; (3) examine whether supematants from activated MBP-specific T cells and PBMCs cause damage and/or death to primary cortical neurons and oligodendrocytes (human), and primary rat oligodendrocytes in culture and whether calpain inhibitors provide neuron protection and restore function. Understanding the mechanisms may help develop therapeutic strategies (e.g., calpain as a target) for treatment of MS.

有力的证据表明，多发性硬化症（MS）是通过自身免疫攻击介导的 髓磷脂特异性T细胞会损害髓鞘的轴突和细胞。了解基础 T细胞被激活，迁移到CM并施加损害的机制对于治疗至关重要 脱髓鞘疾病。 Calpain是一种钙（CA）活化的中性蛋白酶，已与 多发性硬化症。这种竞争的更新应用将决定钙蛋白酶活性的变化是否复发 MS患者和缓解MS患者参与T细胞激活和迁移，TH1/TH2失调，表位 扩散和/或轴突损伤以及神经元和神经胶质细胞死亡，因此检查 对这些事件的抑制作用。初步数据表明钙蛋白酶的表达和活性， 在MS等离子体中，与Interieukin（IL）-2生产（TH1）的伴随着同时增加 与对照组相比，MS患者的外周血单核细胞（PBMC）上清液。这 复发和缓解组的IL-2产生增加与增加的易位 核因子NFATD。相比之下，IL-4和IL-10（TH2）水平比较MS的缓解水平最高 复发。从MS患者细胞释放的Calpain降解MBP和CALPAIN的活性增加了 用干扰素（IFN）-y（MS中增加）处理的原代星形胶质细胞和小胶质细胞。 我们假设MS PBMC和MBP特异性T中的加仑蛋白表达和活性增加 复发和缓解患者的细胞将与Th1/Th2细胞因子谱的变化相关，NFAT 易位和IL-2产生；这释放了Calpain和其他炎症因素 活化的T细胞会损害轴突和神经元，并将MBP降解为抗原肽。钙蛋白酶 抑制剂治疗将抑制T细胞的激活和迁移，减少促炎反应，并 减弱细胞死亡和轴突损伤。这些假设将通过以下特定目的测试：（1） 检查钙蛋白酶表达与活性以及MS中Th1/Th2细胞因子水平之间的相关性 与对照组相比，PBMC和MBP特异性T细胞以及血浆； （2）确定机制 MS PBMC和MBP特异性T细胞的激活和迁移并检查钙蛋白酶的影响 抑制;确定MBP对免疫原性肽的消化是否依赖于CALPAIN，以及是否存在 肽充分激活MS T细胞； （3）检查是否活化的MBP特异性 T细胞和PBMC对原发性皮质神经元和少突胶质细胞造成损害和/或死亡 （人）和原发性大鼠培养物中的少突胶质细胞以及钙蛋白酶抑制剂是否提供神经元 保护和还原功能。了解机制可能有助于制定治疗策略 （例如，钙蛋白酶作为目标）用于治疗MS。

项目成果

Inhibition of calpain attenuates encephalitogenicity of MBP-specific T cells.

• DOI: 10.1111/j.1471-4159.2009.06287.x
• 发表时间: 2009-09
• 期刊: Journal of neurochemistry
• 影响因子: 4.7
• 作者: Guyton MK;Brahmachari S;Das A;Samantaray S;Inoue J;Azuma M;Ray SK;Banik NL
• 通讯作者: Banik NL

Neuron-microglia interaction induced bi-directional cytotoxicity associated with calpain activation.

• DOI: 10.1111/jnc.13774
• 发表时间: 2016-11
• 期刊: Journal of neurochemistry
• 影响因子: 4.7
• 作者: Podbielska M;Das A;Smith AW;Chauhan A;Ray SK;Inoue J;Azuma M;Nozaki K;Hogan EL;Banik NL
• 通讯作者: Banik NL

Regulation of Th1/Th17 cytokines and IDO gene expression by inhibition of calpain in PBMCs from MS patients.

• DOI: 10.1016/j.jneuroim.2010.09.030
• 发表时间: 2011-03
• 期刊: Journal of neuroimmunology
• 影响因子: 3.3
• 作者: Smith AW;Doonan BP;Tyor WR;Abou-Fayssal N;Haque A;Banik NL
• 通讯作者: Banik NL