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: 8066303
• 负责人: Weihua Zhao
• 金额: $ 7.55万
• 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 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8066303
• 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（由申请人提供）：CD 4 + CD 25 + Treg细胞调节小胶质细胞并在ALS中提供神经保护肌萎缩性侧索硬化症（ALS）是最常见的运动神经元疾病，其特征在于脑和脊髓中运动神经元的广泛丧失。ALS的病因不明，也没有已知的治疗方法。以活化的小胶质细胞和浸润的免疫细胞为特征的神经炎症是ALS患者和ALS小鼠的病理标志。目前的证据表明，小胶质细胞/巨噬细胞活化可能是一把双刃剑。许多研究已经得出结论，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。M1/M2巨噬细胞表型已被证明受到CD 4 + T细胞（尤其是CD 4 + CD 25 + T调节（Treg）细胞）的调节。然而，关于ALS中小胶质细胞/单核细胞活化状态与CD 4 + CD 25 + Treg细胞之间的相互作用知之甚少。我们对mSOD 1G 93 A小鼠的初步研究表明，当疾病缓慢进展时，小胶质细胞可能在早期阶段表现出M2表型，而当疾病快速进展时，小胶质细胞可能在晚期阶段表现出M1表型。我们的体内数据显示，缺乏CD 4 + T细胞加速了mSOD 1小鼠的疾病进展。在mSOD 1G 93 A/CD 4-/-小鼠的脊髓中，M2标记物Ym 1以及神经营养因子的表达降低，而炎症因子显著增加。我们确定，当与mSOD 1G 93 A CD 4 + CD 25- T-细胞相比时，mSOD 1G 93 A CD 4 + CD 25 + Treg细胞抑制成年mSOD 1G 93 A小胶质细胞的活化，如通过N 0X 2表达所测量的。此外，CD 4 + CD 25 + Foxp 3 + Treg细胞在疾病的较慢疾病进展阶段扩增。因此，我们的假设是，CD 4 + CD 25 + Treg细胞发挥调节作用，延迟小胶质细胞/单核细胞随着疾病进展从保护性（M2）状态转变为毒性（M1）状态，并且CD 4 + CD 25 + Treg细胞的过继转移将延长mSOD 1转基因小鼠的保护性M2状态并延迟疾病进展。因此，特异性目的1将通过定量RT-PCR、流式细胞术和免疫组织化学使用M1和M2标记物鉴定mSOD 1G 93 A小鼠中不同疾病阶段小胶质细胞/单核细胞的不同活化状态。具体目标2将在体外检查不同小胶质细胞/单核细胞活化状态与CD 4 + CD 25 + Treg细胞之间的相互作用。此外，将在Treg缺失的mSOD 1G 93 A小鼠的脊髓和血液中体内检测M1和M2表型。具体目标3将确定CD 4 + CD 25 + Treg细胞的过继转移是否会延长M2小胶质细胞表型并对mSOD 1G 93 A小鼠产生有益影响。将在转移的小鼠中检查疾病进展、运动神经元损失、小胶质细胞/单核细胞的活化状态。该项目将提供体外和体内证据，证明CD 4 + CD 25 + Treg细胞调节M2小胶质细胞/单核细胞，并在ALS小鼠中提供神经保护。由于ALS患者仅在疾病发作后寻求医疗关注，因此迫切需要针对减缓疾病进展的治疗，例如利用CD 4 + CD 25 + Treg细胞。 公共卫生相关性：ALS是一种可怕的，毁灭性的神经退行性疾病，患者在很短的时间内看着自己恶化，尽管进行了广泛的基础研究，但有效的治疗方法很少。我们自己开发有意义的疗法的努力集中在先天和适应性免疫系统的作用上。最近，T细胞已被证明具有改善ALS模型中神经功能和预期寿命的能力-由于T细胞在ALS患者中很容易获得，因此定义ALS模型中介导神经保护的特定群体可转化为我们在ALS患者中使用T细胞疗法以减缓疾病进展并改善其生活质量的最终目标。