CD4+CD25+ Treg cells regulate microglia and provide neuroprotection in ALS

CD4 CD25 Treg 细胞调节小胶质细胞并在 ALS 中提供神经保护

• 批准号: 7875119
• 负责人: Weihua Zhao
• 金额: $ 7.7万
• 依托单位: METHODIST HOSPITAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7875119
• 关键词: Adoptive Transfer; Adult; Amyotrophic Lateral Sclerosis; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Blood; Brain; CD4 Positive T Lymphocytes; Categories; Cell Therapy; Cells; Coculture Techniques; Data; Disease; Disease Progression; Exhibits; Flow Cytometry; Genes; Goals; Human; IL2RA gene; Immune; Immune system; Immunohistochemistry; Immunosuppressive Agents; In Vitro; Inflammatory; Investigation; Life Expectancy; Location; Lymphocyte; Macrophage Activation; Measures; Mediating; Microglia; Modeling; Motor Neuron Disease; Motor Neurons; Multiple Sclerosis; Mus; Nervous System Physiology; Neurodegenerative Disorders; Onset of illness; Parkinson Disease; Patients; Phase; Phenotype; Play; Population; Process; Production; Quality of life; Reactive Oxygen Species; Regulatory T-Lymphocyte; Reporting; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Spinal Cord; Spinal Cord Diseases; Staging; T-Lymphocyte; Testing; Time; Tissues; Transgenic Mice; Umbilical Cord Blood; Wound Healing; cytokine; effective therapy; end stage disease; improved; in vivo; macrophage; medical attention; monocyte; neuroinflammation; neuroprotection; neurotoxic; neurotrophic factor; overexpression; public health relevance

DESCRIPTION (provided by applicant): CD4+CD25+ Treg cells regulate microglia and provide neuroprotection in ALS Amyotrophic lateral sclerosis (ALS), the most common motor neuron disease, is characterized by the extensive loss of motoneurons in the brain and spinal cord. The cause of ALS is unknown, and there is no known cure. Neuroinflammation, characterized by activated microglia and infiltrating immune cells, is a pathological hallmark in both ALS patients and ALS mice. Current evidence suggests that microglial/macrophage activation may be a double-edged sword. Numerous studies have concluded that alternatively activated macrophages (M2) are protective, while classically activated macrophages (M1) are toxic. The M1/M2 macrophages phenotypes have been shown to be modulated by CD4+ T-cells, especially CD4+CD25+ T regulatory (Treg) cells. However, little is known about the interaction between microglia/monocyte activation states and CD4+CD25+ Treg cells in ALS. Our preliminary studies with mSOD1G93A mice demonstrated that microglia may exhibit an M2 phenotype at early phase when disease was slowly progressing and an M1 phenotype at end stage when disease was rapidly progressing. Our in vivo data showed that the absence of CD4+ T-cells accelerates disease progression of mSOD1 mice. In the spinal cords of mSOD1G93A/CD4-/- mice, expression of an M2 marker, Ym1, as well as neurotrophic factors were decreased, while inflammatory factors were significantly increased. We determined that mSOD1G93A CD4+CD25+ Treg cells inhibited activation of adult mSOD1G93A microglia, as measured by NOX2 expression, when compared with mSOD1G93A CD4+CD25- T- cells. Moreover, CD4+CD25+Foxp3+ Treg cells expanded during slower disease progression phase of disease. Therefore, our hypothesis is that CD4+CD25+ Treg cells play a regulatory role delaying the microglia/monocyte shift from a protective (M2) to a toxic (M1) state as disease progresses and that the adoptive transfer of CD4+CD25+ Treg cells will prolong the protective M2 state and delay disease progression of mSOD1 transgenic mice. Thus, Specific Aim 1 will identify distinct activation states of microglia/monocytes at different disease phases in mSOD1G93A mice by quantitative RT-PCR, flow cytometry and immunohistochemistry using M1 and M2 markers. Specific Aim 2 will examine the interaction between different microglia/monocyte activation states and CD4+CD25+ Treg cells in vitro. Additionally, M1 and M2 phenotypes will be detected in the spinal cords and blood of Treg-depleted mSOD1G93A mice in vivo. Specific Aim 3 will determine if the adoptive transfer of CD4+CD25+ Treg cells prolongs the M2 microglial phenotype and has beneficial effects in mSOD1G93A mice. Disease progression, motoneuron loss, activation states of microglia/monocytes will be examined in the transferred mice. This project will provide in vitro and in vivo evidence that CD4+CD25+ Treg cells regulate M2 microglia/monocytes and provide neuroprotection in ALS mice. Since ALS patients seek medical attention only after disease onset, therapies directed at slowing disease progression, such as utilizing CD4+ CD25+ Treg cells, are critically needed. PUBLIC HEALTH RELEVANCE: ALS is a horrific, devastating neurodegenerative disease in which patients watch themselves deteriorate over a very short period of time, and despite extensive basic investigations, there is minimal effective therapy. Our own efforts to develop meaningful therapies have focused upon the roles of the innate and adaptive immune systems. Recently, T cells have been shown to have the ability to improve neurological function and life expectancy in ALS models - since T cells are readily accessible in ALS patients, defining the specific populations mediating neuroprotection in the ALS models is translatable into our ultimate goal of using T cell therapies in ALS patients to slow disease progression and improve their quality of life.

描述(申请人提供)：在ALS肌萎缩侧索硬化症(ALS)中，CD4+CD25+Treg细胞调节小胶质细胞并提供神经保护，ALS是最常见的运动神经元疾病，其特征是大脑和脊髓中广泛的运动神经元丢失。肌萎缩侧索硬化症的病因尚不清楚，也没有已知的治愈方法。神经炎症是ALS患者和ALS小鼠的一个病理特征，其特征是小胶质细胞激活和免疫细胞渗透。目前的证据表明，小胶质细胞/巨噬细胞的激活可能是一把双刃剑。许多研究得出结论，交替激活的巨噬细胞(M2)具有保护作用，而经典激活的巨噬细胞(M1)具有毒性。巨噬细胞M1/M2表型受CD4+T细胞，尤其是CD4+CD25+T调节(Treg)细胞的调节。然而，在ALS中，小胶质细胞/单核细胞激活状态与CD4+CD25+Treg细胞之间的相互作用还知之甚少。我们对mSOD1G93A小鼠的初步研究表明，在疾病缓慢发展的早期，小胶质细胞可能表现为M2表型，在疾病快速发展的末期，小胶质细胞可能表现为M1表型。我们的体内数据显示，CD4+T细胞的缺失加速了mSOD1小鼠的疾病进展。在mSOD1G93A/CD4-/-小鼠的脊髓中，M2标志物YM1和神经营养因子的表达减少，而炎症因子的表达显著增加。我们确定，与mSOD1G93A CD4+CD25-T细胞相比，mSOD1G93A CD4+CD25+Treg细胞抑制了成年mSOD1G93A小胶质细胞的激活，通过NOX2表达来衡量。此外，CD4+CD25+Foxp3+Treg细胞在疾病进展较慢的阶段扩增。因此，我们的假设是，随着疾病的进展，CD4+CD25+Treg细胞在小胶质细胞/单核细胞从保护性(M2)向毒性(M1)状态的转变中起着调节作用，过继转移CD4+CD25+Treg细胞将延长mSOD1转基因小鼠的保护性M2状态，并延缓疾病的进展。因此，特异性目标1将通过定量RT-PCR、流式细胞术和使用M1和M2标记的免疫组织化学来鉴定mSOD1G93A小鼠不同疾病阶段小胶质细胞/单核细胞的不同激活状态。特殊目的2将在体外检测不同的小胶质细胞/单核细胞激活状态与CD4+CD25+Treg细胞之间的相互作用。此外，在体内Treg缺失的mSOD1G93A小鼠的脊髓和血液中将检测到M1和M2表型。具体目标3将确定过继转移CD4+CD25+Treg细胞是否延长了M2小胶质细胞的表型，并对mSOD1G93A小鼠产生了有益的影响。在转移的小鼠中，将检测疾病进展、运动神经元丢失、小胶质细胞/单核细胞的激活状态。该项目将在体外和体内提供证据，证明CD4+CD25+Treg细胞调节M2小胶质细胞/单核细胞，并为ALS小鼠提供神经保护。由于ALS患者只有在发病后才寻求医疗护理，因此迫切需要针对减缓疾病进展的治疗，如利用CD4+CD25+Treg细胞。 与公共卫生相关：ALS是一种可怕的、破坏性的神经退行性疾病，患者眼睁睁地看着自己在很短的时间内恶化，尽管进行了广泛的基础调查，但有效的治疗方法很少。我们自己努力开发有意义的治疗方法，重点放在先天免疫系统和适应性免疫系统的作用上。最近，在ALS模型中，T细胞被证明具有改善神经功能和预期寿命的能力--由于ALS患者很容易获得T细胞，因此在ALS模型中定义介导神经保护的特定人群可以转化为我们的最终目标，即在ALS患者中使用T细胞疗法来减缓疾病进展并提高他们的生活质量。