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

爱泼斯坦-巴尔病毒(EBV)是疱疹病毒家族的一员，它潜伏地感染B细胞，并已知会导致单核细胞增多症、淋巴癌和上皮癌。流行病学研究表明，EBV感染是多发性硬化症(MS)发展的必经步骤，但其在病理中的作用尚不清楚，自身抗原模拟物也是不可复制的。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自身免疫触发因素和可能在这种免疫反应背后的相应的人类编码模拟抗原。在这里，我们建议从EBV阳性的MS患者的外周血单核细胞(PBMC)中产生重组人抗体库，并利用核糖体展示技术从EBV显示的重组多肽库和健康/疾病的人脑转录文库中进行筛选。因此，这种新的方法将是在一个完全基于体外的选择平台上使用文库来以一种公正和详尽的方式鉴定同源抗体-抗原配对。一旦我们确定了抗EBV抗原抗体和人类开放阅读框架(ORF)抗体，这些抗体也将被用于PAN随机的、通常为12聚体的多肽库，以产生一致的结合基序。然后，这些一致的模体将被用于搜索预测的人类ORF，以确定可能参与EBV结合的候选分子模拟物以及与人类ORF的潜在交叉反应，以确定MS或潜在的其他自身免疫性疾病的靶点。该方法可以应用于该小组的其他研究内容，该项目为学生在博士课程中进一步发展的研究提供了基础。

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.