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: 9087101
• 负责人: William M Ridgway
• 金额: $ 19.75万
• 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/9087101
• 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; 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; 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，显示通过用激动性TLR 4/MD-2特异性单克隆抗体（UT 18，下文称为“TLR 4-Ab”）治疗逆转非肥胖糖尿病（NOD）小鼠中新发1型糖尿病（T1 D）（高血糖症、多尿和体重减轻）。用TLR 4-Ab治疗的小鼠中有90%显示出临床反应（延迟进展至终末期T1 D），70%具有T1 D的永久逆转。成功治疗的小鼠表现出胰岛炎症减少和胰岛β细胞的胰岛素染色保留。我们的方法是高度创新的，因为它靶向先天免疫系统中的特定分子复合物TLR 4/MD-2，以恢复适应性免疫系统的耐受性。虽然TLR 4-Ab不直接刺激T细胞，但我们表明，免疫耐受可以通过这种治疗恢复到适应性免疫系统：来自新发糖尿病小鼠的T细胞，然后用TLR 4-Ab成功治疗两次，不会转移疾病。这种方法对T1 D和其他自身免疫性疾病具有广泛的意义：与小鼠相比，TLR 4/MD-2途径在人类中具有很强的进化保守性，这为在人类中测试这种方法/免疫途径提供了理论基础。此外，已经有FDA批准的靶向人类TLR 4/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，这一途径是一个有前途的新的治疗方法，在自身免疫性测试。我们建议研究逆转T1 D和恢复适应性免疫耐受的免疫学机制，通过这种新的方法实现两个目标。在目标一，“定义TLR 4-Ab治疗的糖尿病小鼠中恢复的适应性免疫T细胞耐受的机制”中，我们将解决令人惊讶的发现，即从已经发展新发糖尿病并成功地用TLR 4-Ab治疗的小鼠中分离的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？是否使CD 4+和CD 8 + T效应细胞（Teff）产生耐受原性或无反应性，或T调节细胞（TcR）的数量或功能增加-或TcR和Teff均受治疗影响？我们将用来自抗体处理的动物的细胞亚型进行研究，以鉴定TLR 4-Ab处理是否调节了T细胞和/或T效应细胞。在目标二，“在T细胞受体转基因转移系统中TLR 4-Ab治疗对T效应细胞和T调节细胞的机械作用的定量和功能分析”中，我们将采用严格定义的T细胞转基因转移系统来定量TLR 4-Ab在自身免疫性糖尿病发作之前、期间和之后对特定细胞亚群的作用，并绘制TLR 4-Ab处理后转基因自身反应性T细胞的命运和功能。了解TLR 4通路介导的疾病逆转的免疫学机制具有巨大的兴趣和潜力。这些目标在两年多的时间里完成，应该明确建立抗TLR 4治疗的疗效和机制，以供将来应用于人类T1 D和其他自身免疫性疾病。