Attenuation of Inflammatory Response in Progressive Neurodegeneration in Parkinson's Disease

帕金森病（PD）是一种进行性退行性疾病，影响着近8万名退伍军人。由于 没有治愈的方法，必须开发新的疗法来阻止疾病的进展。虽然这种退化的机制- 其发生过程尚不清楚，可能涉及慢性炎症。钙蛋白酶激活小胶质细胞和T细胞， 诱导T细胞迁移/趋化性，在脊髓（SC）变性中起关键作用，可能是一种驱动因素， 炎症和疾病进展的迹象。MPTP小鼠和人类PD样本的初步数据显示， CD 4+和CD 8 + T细胞浸润到SC和SN中，增加小鼠脾细胞中的CD 4 + T细胞，以及 血清细胞因子/趋化因子升高。细胞毒性CD 4 + T细胞亚群扩增， 来自MPTP小鼠和DSP-4/6 OHDA诱导的大鼠的脾细胞。钙蛋白酶（钙蛋白酶抑制剂）治疗 在MPTP小鼠中消除了这种CD 4+亚型，表明钙蛋白酶在T细胞活化和CD 4+亚型中的作用。 在炎症过程中可能至关重要趋化因子受体CCR-1是Ca 2+动员剂，重要的是， 可以激活钙蛋白酶由于CCR-1配体（MIP-1α，RANTES）促进SC中T细胞的运输，因此它们在免疫调节中的作用可能与其在免疫调节中的作用有关。 评估炎症。在用钙蛋白酶肽处理MPTP小鼠后，它们显著降低， 行为功能明显改善。PD患者的SC显示小胶质细胞活化， 星形胶质细胞、CD 4 +/CD 8 + T细胞浸润和钙蛋白酶增加。抑制原发性小胶质细胞活化 钙蛋白酶肽和CCR-1拮抗剂（BX 471）的Ca 2+离子载体使细胞因子/细胞周期显著减少。 趋化因子，表明其作为治疗MPTP诱导的神经毒性的药剂的潜力。因此我们 假设钙蛋白酶激活，炎性T细胞（Th 1/Th 17，CD 8+）浸润， 细胞因子/趋化因子参与PD的进行性变性，钙蛋白酶抑制剂和CCR-1 拮抗剂治疗可以减少变性、减缓疾病进展和改善功能。三 具体目标是：（1）研究钙蛋白酶调节和T细胞浸润在SC退变中的作用， 在MPTP小鼠中的免疫应答和疾病进展，表征浸润性T细胞，评估细胞因子/趋化因子水平 在血清中，并测定SC中的细胞死亡参数和钙蛋白酶活化;（2）检查T细胞的浸润， PD患者SC中小胶质细胞的活化，表征T细胞亚群，并与特异性 目的：1、检测PD患者血清中趋化因子/细胞因子水平; 单独或联合使用钙蛋白酶抑制剂（钙蛋白酶，SNJ 1945）和BX 471的MPTP小鼠将减少炎症反应。 炎症事件和退化和改善功能，与DigiGait分析相关，并检查是否 由钙离子载体激活的初级小胶质细胞上调趋化因子/细胞因子， 钙蛋白酶肽和BX 471。MPTP小鼠中CD 4亚型的关键发现也将在鱼藤酮中得到验证， DSP-4/6 OHDA大鼠模型。其目标是开发PD治疗策略，使用阻断炎症的药物， 炎症过程，保护神经元，控制疾病进展，改善功能。

Parkinson's disease (PD), a progressive degenerative disorder, affects almost 80,000 Veterans. Since there is no cure, new therapies must be developed to halt disease progression. While the mechanisms of this degen- erative process remain elusive, chronic inflammation may be involved. Calpain activates microglia and T cells, induces T cell migration/chemotaxis, plays a pivotal role in spinal cord (SC) degeneration, and may be a driver of inflammation and disease progression. Preliminary data from MPTP mice and human PD samples showed infiltration of CD4+ and CD8+ T cells into SC and SN, increased CD4+ T cells in mouse splenocytes, and elevated serum cytokines/chemokines. An expanded subpopulation of cytotoxic CD4+ T cells was detected in splenocytes from MPTP mice and DSP-4/6OHDA-induced rats. Calpeptin (calpain inhibitor) treatment abolished this CD4+ subtype in MPTP mice, suggesting calpain's role in T cell activation and the CD4+ subtype may be critical in the inflammatory process. Chemokine receptor CCR-1 is a Ca2+ mobilizer and, importantly, can activate calpain. Since CCR-1 ligands (MIP-1α, RANTES) promote trafficking of T cells in SC, their role in inflammation was assessed. They were significantly reduced following treatment of MPTP mice with calpeptin, and behavioral function was remarkably improved. SC from PD patients revealed activation of microglia and astrocytes, infiltration of CD4+/CD8+ T cells, and increased calpain. Inhibition of primary microglia activation by Ca2+ ionophore by calpeptin and CCR-1 antagonist (BX471) produced marked reduction in cytokines/ chemokines, suggesting their potential as agents for treatment of MPTP-induced neurotoxicity. Thus, we hypothesize that calpain activation, infiltration of inflammatory T cells (Th1/Th17, CD8+), and released cytokines/chemokines are involved in progressive degeneration in PD, and calpain inhibitor and CCR-1 antagonist treatment may reduce degeneration, slow disease progression, and improve function. Three specific aims are proposed: (1) investigate the role of calpain regulation and T cell infiltration in SC degenera- tion and disease progression in MPTP mice, characterize infiltrating T cells, assess cytokine/chemokine levels in sera, and determine cell death parameters and calpain activation in SC; (2) examine infiltration of T cells and activation of microglia in SC from PD patients, characterize T cell subpopulations, and correlate with Specific Aim 1, determine chemokines/cytokines in serum from PD patients; and (3) examine whether treatment of MPTP mice with calpain inhibitors (calpeptin, SNJ1945) and BX471 alone or in combination will reduce inflam- matory events and degeneration and improve function, correlate with DigiGait analysis, and examine whether primary microglia activated by calcium ionophore upregulate chemokines/cytokines and can be controlled by calpeptin and BX471. The key finding of a CD4-subtype in MPTP mice will also be verified in rotenone and DSP-4/6OHDA rat models. The goal is to develop strategies for PD therapy with agents that block the inflam- matory process, protect neurons, control disease progression, and improve function.