Targeting GSK-3 in antigen-specific immunotherapy

1911年，努恩和弗里曼发表了他们用花粉提取物治疗花粉热患者的报告。这是最早记录的特异性免疫疗法（SIT）的描述之一，其中使用引起过敏状况的药物来“脱敏”患者。虽然SIT的价值已被广泛接受，但该方法仍然存在严重副作用的风险，包括过敏性休克。在自身免疫性疾病治疗方面，SIT的研究落后于其在过敏方面的研究，并且还受到异常事件的阻碍。例如，在灵长类动物中，给药一种脑蛋白会加重一种类似多发性硬化症的疾病。同样，在动物模型中，通过肺或肠道给药人工胰岛抗原诱导T淋巴细胞，加重自身免疫性糖尿病。与完整抗原相关的并发症导致该领域的意见领袖建议使用称为“合成肽”的蛋白质片段用于SIT。我们的实验室研究设计和肽的作用模式，能够抑制免疫反应。抑制的有效诱导取决于肽的溶解度。以前我们已经表明，即使以高可溶性形式施用，一些肽也不能介导SIT。事实证明，这是因为肽可以与它们的受体——一种叫做MHC的免疫分子结合，从而产生错误的形状或构象。然而，多肽可以被设计成适合MHC，然后诱导对天然蛋白质的耐受性。有效的SIT与多肽会诱导淋巴细胞亚群，即CD4 t细胞，它们分泌一种叫做IL-10的蛋白质。IL-10是一种激素样物质，被称为细胞因子，通过负反馈循环机制抑制体内邻近T细胞的激活。最近的研究表明，过敏性哮喘中的肽SIT也产生IL-10依赖性抑制，从而揭示了IL-10在过敏和自身免疫性疾病中的SIT中的类似作用。目前这项提议的总体目标是通过增强能够引起疾病的淋巴细胞向分泌IL-10的疾病抑制细胞的转化来提高肽诱导的SIT的疗效。最近我们发现，在体外培养的淋巴细胞中加入阻断GSK-3酶活性的药物可大大提高其IL-10的产生。因此，该项目的具体目标是：1。从先前由GSK ii表征的面板中确定最合适的GSK-3抑制剂。在相关的自身免疫性疾病动物模型中，研究GSK-3抑制剂与SIT联合促进体内IL-10分泌抑制细胞生成的能力ii。确认所选抑制剂对人淋巴细胞的活性。评估小鼠和人类细胞中IL-10表型的稳定性这些化合物没有专利，因此，不存在与它们相关的“操作自由”问题。我们提出了这些化合物作为辅助治疗的新应用，与SIT结合，用于自身免疫性疾病和过敏的免疫治疗。我们进行了与该药物申请相关的专利检索，未发现现有技术。在SIT期间IL-10产生的增强将大大提高这种治疗方法的疗效。因此，GSK-3抑制剂作为辅助治疗将适用于目前使用SIT治疗的任何过敏性疾病，或同样适用于已知目标抗原的任何自身免疫性疾病。共同施用GSK-3抑制剂所提供的SIT改善将使医疗保健提供者不再使用非特异性治疗过敏和自身免疫性疾病，因为这些治疗通常无效且可能导致严重的副作用。这一改进将对患者和卫生保健系统都有相当大的好处。

In 1911, Noon and Freeman published their account of the treatment of a hay fever sufferer with pollen extract. This was one of the first documented descriptions of specific immunotherapy (SIT) in which the agent causing the allergic condition was used to 'desensitise' the patient. While the value of SIT has been widely accepted, the approach still carries the risk of severe side effects including anaphylactic shock. The investigation of SIT for treatment of autoimmune diseases has lagged behind that of SIT in allergy and has also been hampered by aberrant events. For example, administration of a brain protein exacerbated a disease resembling MS in primates. Similarly, administration of an artificial islet antigen, via the lung or gut, induced T lymphocytes that exacerbated autoimmune diabetes in an animal model. The complications associated with intact antigen have led opinion leaders in the field to recommend the use of fragments of proteins called 'synthetic peptides' for SIT. Our laboratory studies the design and mode of action of peptides that are able to suppress immune responses. Effective induction of suppression depends on the solubility of the peptide. Previously we had shown that some peptides failed to mediate SIT even when administered in a highly soluble form. This proved to be because the peptides could bind to their receptor, an immune molecule called MHC, to produce the wrong shape or conformation. Peptides can be designed, however, to fit the MHC appropriately and these then induce tolerance to the native protein. Effective SIT with peptides leads to the induction of a subset of lymphocytes, known as CD4 T-cells, and they secrete a protein called IL-10. IL-10 is a hormone-like substance, known as a cytokine, that suppresses activation of neighbouring T cells in the body via a negative feedback loop mechanism. It was recently shown that peptide SIT in allergic asthma also generates IL-10-dependent suppression thus revealing a similar role for IL-10 in SIT for both allergy and autoimmune disease.The overall aim of this current proposal is to improve the efficacy of peptide-induced SIT by enhancing the conversion of lymphocytes capable of causing disease into IL-10 secreting disease suppressor cells. Recently we showed that the addition of drugs that block the activity of the enzyme GSK-3 to cultures of lymphocytes in vitro greatly enhanced their production of IL-10. The specific aims of this project are, therefore, to:i. identify the most suitable GSK-3 inhibitor from a panel previously characterised by GSK ii. investigate the ability of GSK-3 inhibitors to enhance the in vivo generation of IL-10 secreting suppressor cells in conjunction with SIT in a relevant animal model of autoimmune diseaseiii. confirm the activity of the selected inhibitor/s on human lymphocytesiv. assess the stability of the IL-10 phenotype in both mouse and human cellsThere are no patents relating to these compounds and, as a result, no 'freedom to operate' issues associated with them. We propose a novel application for these compounds, as adjunct treatments, in conjunction with SIT, for immunotherapy of autoimmune diseases and allergies. We have conducted patent searches relating to this application of the drugs and have not found prior art. The enhancement of IL-10 production during SIT will greatly improve the efficacy of this therapeutic approach. As such, the use of GSK-3 inhibitors as adjunct therapy will be applicable to any allergic condition currently treated by SIT or equally to any autoimmune condition for which the target antigens are known. The improvement of SIT provided by co-administration of GSK-3 inhibitors will allow health care providers to move away from the use of non-specific therapies for allergic and autoimmune diseases since these are often ineffective and can cause severe side effects. This improvement will be of considerable benefit to both patients and the health care system.