Studies on the aetiology/mechanisms of autoimmunity associated with Epstein Barr Virus infection

爱泼斯坦巴尔病毒（EBV）是疱疹病毒家族的成员，其潜伏感染B细胞，并且已知引起单核细胞增多症以及淋巴和上皮癌。流行病学研究表明，EB病毒感染是多发性硬化症（MS）发展的必然步骤，但其在病理学中的作用仍不清楚，并且自身抗原模拟物无法重现。自身免疫的发展是多基因和环境影响的产物，EBV感染诱导的免疫反应被认为可能是MS的驱动力。我们的目的是阐明关键的EBV自身免疫触发因子和相应的人类编码的模拟抗原，可能是这种免疫反应的背后。在这里，我们建议从外周血单个核细胞（PBMC）的EBV阳性的人MS产生重组人抗体库，并选择对重组EBV展示的多肽库和对健康/患病的人脑转录组库使用核糖体展示技术。因此，这种新方法将在完全基于体外的选择平台中“使用文库对抗文库”，以无偏倚和穷举的方式鉴定同源抗体-抗原配对。一旦我们鉴定了抗EBV抗原抗体和人开放阅读框（ORF）抗体，这些抗体也将用于淘选随机的、通常为12聚体的肽文库以产生共有结合基序。然后将使用共有基序来搜索预测的人ORF，以鉴定参与EBV结合和与人ORF的潜在交叉反应性的可能的候选分子模拟物，以鉴定MS或潜在的其他自身免疫性疾病的靶标。该方法可以应用于其他研究元素与组和该项目提供了一个研究的基础，可以由学生在博士学位的过程中进一步发展。

Epstein Barr Virus (EBV) is a member of the herpesvirus family that latently infects B cells and is known to cause mononucleosis and lymphoid and epithelial cancer. Epidemiological studies suggest EBV infection is an obligate step for the development of multiple sclerosis (MS), but its role in pathology remains unclear, and autoantigen mimics have been unreproducible. Development of autoimmunity is the product of polygenic and environmental influences, and infection with EBV induces an immune response that is believed to possibly be the driving force of MS. Our aim is to elucidate the key EBV autoimmune triggers and the corresponding human encoded mimetic antigens that could be behind this immune response. Here, we propose to generate recombinant human antibody libraries from the peripheral blood mononuclear cells (PBMC) from EBV-positive people with MS and to select against a recombinant EBV-displayed polypeptide library and against healthy/diseased human brain transcriptomic libraries using ribosome display technology. This novel approach will therefore be "using a library against a library" in a completely in-vitro-based selection platform to identify the cognate antibody-antigen pairings in an unbiased and exhaustive way. Once we have identified the anti-EBV antigen antibodies and human open reading frame (ORF) antibodies, these will also be used to pan a random, typically 12mer peptide library to generate consensus binding motifs. The consensus motifs will then be used to search the predicted human ORFs to identify likely candidate molecular mimics involved in EBV-binding and potential cross-reactivity with human ORFs, to identify targets for MS or potentially other autoimmune diseases. The approach could be applied to other research elements with the group and the project provides a basis for studies that can be developed further by the student over the course of a PhD.