Characterisation of autoantigen-specific human B cells in central nervous system autoantibody-mediated diseases

背景我们的免疫系统已经进化到保护我们免受感染。通常，最有效的保护是通过其产生抗体。抗体由免疫系统的“B细胞”产生。抗体可以与细菌结合并直接清除感染。然而，有时候，他们会错误地将我们自己的身体视为入侵者。这些所谓的“自身抗体”导致“自身免疫”疾病，影响约10%的人口，并导致严重残疾，有时甚至死亡。因此，了解身体如何去除自我反应的B细胞对于开发更好的治疗方法至关重要。自身免疫性疾病几乎可以影响身体的任何组织，但传统上，大脑被认为相对受到保护。然而，在过去的十年中，我们和其他人已经成功地鉴定了15种被自身抗体靶向的脑蛋白。这远远超过了其他任何分支医学中类似的自身免疫综合征的数量。患有这些自身抗体的患者通常会出现癫痫发作、记忆丧失和精神症状，统称为“脑炎”。自身免疫性脑炎（AE）的数量每年持续增长。牛津大学是英国AE患者的主要转诊中心。95%的患者同意参与研究，因此我的团队可以直接研究患者样本，并将结果与他们的症状和治疗反应联系起来。事实上，我们的患者在使用抑制其免疫系统的药物后表现出一些改善。然而，80%的人无法重返工作岗位，复发或经历药物治疗不可接受的副作用。为了改善患者的这些结果，该项目旨在指导发现改进的药物。此外，由于自身免疫性疾病是常见的，B细胞在许多神经系统疾病的大脑中被发现，我们的研究结果将为其他几种疾病患者的生物学和护理提供信息。确定哪种B细胞最先发生自我反应，哪种细胞的自我反应最强。这将突出未来治疗的理想目标细胞，为目标2和目标3奠定基础，并为我们的患者提供因果关系的解释。这项工作将研究我们的患者和健康人捐献的血液。我们将血液B细胞分成代表其发育阶段的群体。然后，可以激活个体群体以发现最早和最有效的自身反应性抗体生产者。与健康人的比较将显示自我反应性B细胞被成功清除的位置。通过在患者接受治疗后重复这些研究，我们将了解目前的疗法是否可以将该过程重置为健康状态下观察到的过程。2.描述仅在产生自身抗体的B细胞上发现的分子。用药物靶向这些分子将同时增加有效性和减少副作用。利用目标1的发现，将纯化血液中最具自身反应性的B细胞群。此外，来自患者脊髓液的样本将接触自身反应性B细胞，这些细胞循环并接触其最终目标-大脑。从这两个网站，我们的目标是比较自我反应的非自我反应的细胞，并确定前者的分子的不同范围。将与牛津大学和耶鲁大学（美国）的细胞分析专家合作开展工作。去询问某些自身抗体是否特别能诱发疾病。制造这些抗体的B细胞将成为未来治疗的更精确靶点。从Aim 2中纯化的B细胞中，我们将回收多种自身抗体。这些将被应用于“培养皿中”的脑细胞，并注射到小鼠体内，以确定那些最有力地破坏这些系统的细胞。将其与Aim 2的数据相关联，将揭示产生最有效自身抗体的B细胞的概况。

BackgroundOur immune system has evolved to protect us from infections. Often, the most effective protection is via its production of antibodies. Antibodies are made by the immune system's 'B cells'. Antibodies can bind to bacteria and directly clear an infection. However, sometimes, they can - in error - recognise our own body as the invader. These so-called 'autoantibodies' lead to 'autoimmune' diseases which affect ~10% of the population and causes significant disability and sometimes death. So, understanding how the body removes the self-reactive B cells is of major importance to develop better treatments.Autoimmune conditions can affect almost any tissue in the body but, traditionally, the brain has been considered relatively protected. However, over the last decade, we and others have successfully identified 15 brain proteins which are targeted by autoantibodies. This far outweighs the number of similar autoimmune syndromes in any other branch of medicine. Patients with these autoantibodies often have seizures, memory loss and psychiatric symptoms, together called 'encephalitis'. The number of forms of autoimmune encephalitis (AE) continues to grow annually.Oxford is the UK's major referral centre for patients with AE. 95% of our patients consent to research involvement, so my team can directly study patient samples and link findings to their symptoms and treatment responses. Indeed, our patients show some improvements with available medications which suppress their immune system. However, 80% either cannot return to work, have relapses or experience unacceptable side effects from their medications.To improve these outcomes for patients, this project aims to direct the discovery of improved medications. Also, as autoimmune diseases are common and B cells are found in the brain in many neurological conditions, our findings will all inform the biology and care of patients with several other illnesses.Aims of this project:1. To identify which B cells are the first to become self-reactive and which are the most self-reactive. This will highlight cells that are ideal targets for future therapies, lay the foundations for Aims 2 and 3, and offer explanations about causation to our patients.This work will study blood donated by our patients and by healthy people. We will separate blood B cells into populations which represent stages of their development. Then, individual populations can be activated to discover the earliest, and the most potent, producers of the autoreactive antibodies. Comparisons to healthy people will show where the self-reactive B cells are successfully cleared. By repeating these studies after patients are treated, we will understand if current therapies can reset the process to that observed in health. 2. To describe molecules exclusively found on the B cells which make the autoantibodies. Targeting these molecules with medications would simultaneously increase effectiveness and reduce side-effect profiles. Using findings from Aim 1, the most self-reactive B cell populations in blood will be purified. Also, samples from patient spinal fluid will access self-reactive B cells which circulate and contact their ultimate target - the brain. From both sites, we aim to compare the self-reactive to the non self-reactive cells and identify the distinct range of molecules on the former. Work will occur in collaboration with expert cell profilers at Oxford University and Yale University (USA).3. To ask if certain autoantibodies are especially potent at inducing disease. The B cells that make these would be even more precise targets for future therapies.From the B cells purified in Aim 2, we will recover multiple autoantibodies. These will be applied to brain cells 'in a dish' and injected into mice, to identify those which most robustly disrupt these systems. Relating this to data from Aim 2 will reveal the profile of the B cells that made the most potent autoantibodies.