Targeting GSK-3 in antigen-specific immunotherapy

1911年，Noon和Freeman发表了他们用花粉提取物治疗花粉热患者的报道。这是第一个记录的描述特异性免疫疗法（SIT），其中引起过敏性疾病的药物被用来“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。SIT治疗自身免疫性疾病的研究已经落后于SIT在变态反应中的研究，并且也受到异常事件的阻碍。例如，给灵长类动物注射一种脑蛋白会加重一种类似多发性硬化症的疾病。类似地，通过肺或肠道给予人工胰岛抗原，诱导T淋巴细胞，从而在动物模型中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推荐使用称为“合成肽”的蛋白质片段进行SIT。我们的实验室研究能够抑制免疫反应的肽的设计和作用模式。抑制的有效诱导取决于肽的溶解度。以前，我们已经表明，一些肽未能介导SIT，即使在高度可溶性的形式给药。这被证明是因为这些肽可以与它们的受体，一种称为MHC的免疫分子结合，产生错误的形状或构象。然而，可以设计肽以适当地适应MHC，然后这些肽诱导对天然蛋白质的耐受性。有效的SIT与肽导致诱导的淋巴细胞亚群，称为CD 4 T细胞，他们分泌一种蛋白质称为IL-10。IL-10是一种细胞因子样物质，称为细胞因子，通过负反馈回路机制抑制体内邻近T细胞的激活。最近的研究表明，肽SIT在过敏性哮喘中也产生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的SIT中的类似作用。目前建议的总体目标是通过增强能够引起疾病的淋巴细胞转化为分泌IL-10的疾病抑制细胞来提高肽诱导的SIT的功效。最近，我们发现，在体外淋巴细胞培养物中加入阻断GSK-3酶活性的药物，大大提高了IL-10的产生。因此，该项目的具体目标是：从先前通过GSK ii表征的组中鉴定最合适的GSK-3抑制剂。研究GSK-3抑制剂在相关的自身免疫性疾病动物模型中结合SIT增强IL-10分泌抑制细胞的体内产生的能力。确认所选抑制剂对人淋巴细胞的活性。评估IL-10表型在小鼠和人细胞中的稳定性没有与这些化合物相关的专利，因此，没有与它们相关的“操作自由”问题。我们提出了一种新的应用这些化合物，作为辅助治疗，结合SIT，免疫治疗自身免疫性疾病和过敏。我们已经进行了与药物的这种应用相关的专利检索，并且没有发现现有技术。在SIT期间IL-10产生的增强将极大地提高这种治疗方法的功效。因此，使用GSK-3抑制剂作为辅助疗法将适用于目前通过SIT治疗的任何过敏性病症或同样适用于靶抗原已知的任何自身免疫性病症。通过共同施用GSK-3抑制剂提供的SIT的改善将允许医疗保健提供者不再使用用于过敏性和自身免疫性疾病的非特异性疗法，因为这些疗法通常是无效的并且可能引起严重的副作用。这项改善对病人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.