Mechanism of restored immune tolerance in anti-TLR4 antibody reversal of NOD T1D

抗TLR4抗体逆转NOD T1D恢复免疫耐受的机制

• 批准号: 8967990
• 负责人: William M Ridgway
• 金额: $ 23.71万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-15 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8967990
• 关键词: Address; Affect; Animals; Antibodies; Antigens; Apoptosis; Autoimmune Diabetes; Autoimmune Diseases; Autoimmunity; Beta Cell; Biological Assay; Biological Preservation; Blood Glucose; Body Weight decreased; CD8B1 gene; Cell Communication; Cells; Child; Clinical; Complex; Data; Development; Diabetes Mellitus; Diabetic mouse; Disease; Drug Targeting; Effector Cell; Eragrostis; FDA approved; Failure; Figs - dietary; Future; Grant; Health Care Costs; Histologic; Human; Hyperglycemia; Immune; Immune Tolerance; Immune system; Inbred NOD Mice; Inflammation; Insulin; Insulin-Dependent Diabetes Mellitus; Length; Maps; Mediating; Modeling; Molecular; Monoclonal Antibodies; Mus; Natural Immunity; Non obese; Output; Pathway interactions; Polyuria; Public Health; Recurrent disease; Regulatory T-Lymphocyte; Research; Sorting - Cell Movement; Staging; Staining method; Stains; Stimulus; System; T-Cell Receptor; T-Lymphocyte; TLR4 gene; Testing; Transgenic Mice; Transgenic Organisms; United States National Institutes of Health; Work; autoreactive T cell; cancer immunotherapy; cell type; clinical remission; diabetic; effective therapy; high reward; high risk; innovation; insulin dependent diabetes mellitus onset; interest; islet; meetings; novel strategies; novel therapeutic intervention; public health relevance; response; restoration

 DESCRIPTION (provided by applicant): We have exciting new data showing reversal of new onset type 1 diabetes (T1D) (hyperglycemia, polyuria and weight loss) in nonobese diabetic (NOD) mice by treatment with an agonistic TLR4/MD-2 specific monoclonal antibody (UT18, hereafter referred to as "TLR4-Ab"). 90% of mice treated with TLR4-Ab showed a clinical response (delay in progression to endstage T1D) and 70% have permanent reversal of T1D. Successfully treated mice demonstrate decreased islet inflammation and preserved insulin staining of islet beta cells. Our approach is highly innovative because it targets a specific molecular complex, TLR4/MD-2, in the innate immune system to restore tolerance in the adaptive immune system. Although TLR4-Ab does not stimulate T cells directly, we show that immune tolerance can be restored to the adaptive immune system by this treatment: T cells from mice that had developed new onset diabetes, and then were successfully treated twice with TLR4-Ab, do not transfer disease. This approach has broad implications for T1D and other autoimmune diseases: the TLR4/MD-2 pathway is strongly evolutionarily conserved in humans compared to mice, providing a rationale to test this approach/immune pathway in humans. Moreover, there are already FDA approved drugs that target TLR4/MD-2 in humans, and more are actively being developed as a novel approach in cancer immunotherapy. We submit that this pathway is a promising new therapeutic approach for testing in autoimmunity. We propose to study the immunological mechanisms of reversal of T1D and restoration of adaptive immune tolerance achieved by this novel approach in two aims. In aim one, "Defining the mechanisms of restored adaptive immune T cell tolerance in diabetic mice treated with TLR4-Ab", we will address the surprising finding that T cells isolated from mice that had developed new onset diabetes and were successfully treated with TLR4-Ab do not transfer diabetes. By what mechanism were these cells rendered unable to cause disease? Were CD4+ and CD8+ T effectors (Teffs) rendered tolerogenic or anergic, or were T regulatory cells (Tregs) increased in quantity or function - or were both Tregs and Teffs affected by treatment? We will perform studies with cell subtypes from antibody treated animals to identify whether Tregs and/or T effector cells are modulated by TLR4-Ab treatment. In Aim Two, "Quantitative and functional analysis of the mechanistic effects of TLR4-Ab treatment on T effector and T regulatory cells in a T cell receptor transgenic transfer system", we will employ a rigorously defined T cell transgenic transfer system to quantify TLR4-Ab effects on specific cell subsets before, during and after onset of autoimmune diabetes, and map the fate and function of transgenic autoreactive T cells after TLR4-Ab treatment. Understanding the immunological mechanisms of disease reversal mediated by the TLR4 pathway is of tremendous interest and potential. These aims performed over two years should definitely establish the efficacy and mechanisms of anti-TLR4 treatment for future application to human T1D and other autoimmune diseases.

 描述（由申请人提供）：我们有令人兴奋的新数据显示，通过激动性 TLR4/MD-2 特异性单克隆抗体（UT18，以下称为“TLR4-Ab”）治疗，非肥胖糖尿病 (NOD) 小鼠新发 1 型糖尿病 (T1D)（高血糖、多尿和体重减轻）得到逆转。 90% 接受 TLR4-Ab 治疗的小鼠表现出临床反应（延迟进展至终末期 T1D），70% 的 T1D 得到永久性逆转。成功治疗的小鼠表现出胰岛炎症减少并保留胰岛β细胞的胰岛素染色。我们的方法具有高度创新性，因为它针对先天免疫系统中的特定分子复合物 TLR4/MD-2，以恢复适应性免疫系统的耐受性。尽管TLR4-Ab不直接刺激T细胞，但我们表明通过这种治疗可以恢复适应性免疫系统的免疫耐受性：来自患有新发糖尿病的小鼠的T细胞，然后用TLR4-Ab成功治疗两次，不会转移疾病。这种方法对 T1D 和其他自身免疫性疾病具有广泛的影响：与小鼠相比，TLR4/MD-2 通路在人类中在进化上高度保守，这为在人类中测试这种方法/免疫通路提供了理论基础。此外，FDA 已经批准了针对人类 TLR4/MD-2 的药物，并且正在积极开发更多药物作为癌症免疫治疗的新方法。我们认为该途径是一种有前途的自身免疫测试新治疗方法。我们提议研究通过这种新方法实现的 T1D 逆转和适应性免疫耐受恢复的免疫学机制，有两个目的。在目标一“确定用 TLR4-Ab 治疗的糖尿病小鼠中恢复适应性免疫 T 细胞耐受性的机制”中，我们将解决一个令人惊讶的发现，即从患有新发糖尿病并成功用 TLR4-Ab 治疗的小鼠中分离出的 T 细胞不会转移糖尿病。通过什么机制使这些细胞无法引起疾病？ CD4+ 和 CD8+ T 效应细胞 (Teff) 是否变得耐受或无反应，或者 T 调节细胞 (Treg) 的数量或功能是否增加，或者 Tregs 和 Teff 是否都受到治疗的影响？我们将对抗体治疗动物的细胞亚型进行研究，以确定 Tregs 和/或 T 效应细胞是否受到 TLR4-Ab 治疗的调节。在目标二“T细胞受体转基因转移系统中TLR4-Ab治疗对T效应细胞和T调节细胞的机制影响的定量和功能分析”中，我们将采用严格定义的T细胞转基因转移系统来量化TLR4-Ab在自身免疫性糖尿病发病之前、期间和之后对特定细胞亚群的影响，并绘制转基因自身反应性T细胞在发病后的命运和功能图谱。 TLR4-抗体治疗。了解 TLR4 通路介导的疾病逆转的免疫机制具有巨大的兴趣和潜力。这些两年多的目标肯定会确立抗 TLR4 治疗的功效和机制，以便将来应用于人类 T1D 和其他自身免疫性疾病。