Molecular understanding of self immunity in the brain

我最近开发了一种可编程噬菌体展示，用于识别与人类副肿瘤性神经系统疾病（PND）相关的致病抗原。这个工作场所我在迅速崛起的神经免疫学领域，作为PND是衰弱的中枢神经系统疾病，涉及自身免疫性脑炎。PND患者体内含有抗脑抗原的自身抗体，自身抗体谱的特异性可预测临床症状和预后。我已经确定了与患者PND特异性相关的最大一组抗原的特征，其中之一KLHL 11是新的，最近发表。在目标1中，我将表征额外的抗原并测试我的假设，即单个表位是PND患者自身免疫和脑炎的原因。K99将使我获得表位分析所需的高级生物信息学技能，这是导师实验室的优势。在目的2中，我测试是否细胞内PND抗原引起自身免疫性脑炎通过非经典的MHC-I类依赖性途径，一个意想不到的机制与治疗的影响。我将创建一个新的小鼠模型抗KLHL 11 PND和互补KLHL 11敲除小鼠。与现有的小鼠免疫基因缺失，我将使用免疫组织化学（IHC）和单细胞测序来测试在PND的非经典MHC途径的大脑中的T细胞的特异性靶向的依赖性。K99将允许我，与课程和顾问一起，获得目标2和3所必需的免疫学先进知识，并帮助指导最初的T细胞研究。此外，加州大学旧金山分校的使命，包括基础设施和研究环境，特别是面向促进基础和临床免疫学家和神经学家之间的直接互动。我计划继续利用这个特殊的环境，因为我过渡到R 00阶段，在这个目标。在目标3中，我利用我之前在小鼠遗传学和细胞神经生物学方面的专业知识以及我的K99获得的分子免疫学专业知识，利用PND抗原（目标1和2）生成大脑中靶向炎症的3D解剖图。我将测试是否大脑区域特异性PND脑炎是由中枢神经系统淋巴系统介导的。在PND开始后的不同时间对T细胞进行脑清除的IHC将提供入侵T细胞及其与抗原表达细胞的关系的全面“3D”路线图。我提出的研究将导致全面了解神经元抗原如何引起脑自身免疫反应的机制，并假设T细胞迁移途径对解剖学上不同的自身免疫是特异性的，为识别不同PND病理基础的神经元电路铺平道路，我作为独立研究者的目标。

I recently developed a programmable phage display for identifying pathogenic antigens related to human paraneoplastic neurological disorders (PNDs). This workplace me in the rapidly emerging area of neuro- immunology, as PNDs are debilitating central nervous system disorders that involve autoimmune encephalitis. Patients with PND harbor autoantibodies against brain antigens, and specificity in the autoantibody profile predicts clinical symptoms and prognosis. I have characterized the largest set of antigens that are linked specifically to patient PNDs, one of which, KLHL11, was novel and published recently. In Aim 1, I will characterize the additional antigens and test my hypothesis that a single epitope is causal to PND patient autoimmunity and encephalitis. The K99 will allow me to acquire the advanced bioinformatics skills necessary for epitope analysis, strengths of the mentor's lab. In Aim 2, I test whether intracellular PND antigens cause autoimmune encephalitis through a non-canonical MHC-I class-dependent pathway, an unexpected mechanism with implications for treatment. I will create a new mouse model for anti-KLHL11 PND and a complementary KLHL11 knockout mouse. Together with existing mice deleted in immunity genes, I will use immunohistochemistry (IHC) and single cell sequencing to test dependence in specific targeting of T-cells in the brain of PND on the non-canonical MHC pathway. The K99 will allow me, with classes and advisors, to acquire the necessary advanced knowledge in immunology built into Aims 2 and 3 and help guide the initial T cell studies. Further, the UCSF mission, including infrastructure and research environment, is geared particularly towards facilitating direct interactions between basic and clinical immunologists and neurologists. I plan to continue to exploit this specialized environment as I transition to the R00 phase in this aim. In Aim 3, I bring to bear my previous expertise in mouse genetics and cellular neurobiology, and my K99 acquired molecular immunology expertise, to leverage PND antigens (Aims 1 and 2) to generate 3D anatomical maps of targeted inflammation in the brain. I will test whether brain region-specific PND encephalitis is mediated by the CNS lymphatic system. IHC with brain clearing for T cells, at different times after initiation of PND, will provide a comprehensive “3D” roadmap of the invading T cells and their relationship to antigen-expressing cells. My proposed studies will result in a comprehensive picture of the mechanisms determining how neuronal antigens elicit a brain autoimmune response, and assuming T cell migration routes are specific for anatomically distinct auto immunities, pave the way for identifying neuronal circuitries underlying the distinct PND pathologies, my goal as an independent investigator.