Repertoire and antigen recognition of brain-infiltrating immune cells in progressive MS

在多发性硬化症(MS)患者的大脑中发现了来自免疫系统的细胞(例如T细胞和B细胞/浆细胞)，这些细胞位于髓鞘被破坏和神经元退化的区域。此外，在疾病的最开始、复发-缓解阶段和疾病的进展阶段，这些细胞都在多发性硬化症的大脑中被发现。这些细胞是属于几个出错的克隆，还是属于许多不同的克隆？它们识别什么？在MS非常早期的大脑中发现的免疫细胞识别的抗原与后来在疾病进展期发现的免疫细胞识别的抗原相同吗？对于进展性MS患者来说，寻找这些问题的答案的研究还没有完成，因为合适的材料极其罕见，而且通常是甲醛固定和石蜡包埋的(FFPE)被认为对进一步分析毫无用处。然而，我们拥有如此珍贵的样本，我们拥有使用FFPE组织进行分子研究的专业知识。对组织内致病淋巴细胞的研究需要像下一代测序这样的复杂技术来更多地了解它们的谱系(也就是了解它们的抗体和T细胞受体有多大不同，以及它们属于几个还是多个不同的克隆)。在过去的几年里，这些技术也被调整到FFPE样品。与疾病相关的细胞通常在组织内扩张。因此，我们接下来将克隆和表达在进展性多发性硬化症大脑中发现的最丰富的抗体和T细胞受体，以找出这些分子识别什么。最后，当我们知道这一点时，我们将回到病理学来找出这种认识发生的时间和地点。这些问题的答案可能有助于更好地了解多发性硬化症的驱动力，甚至可能找到更好的治疗多发性硬化症的方法。不难想象，只有所有这些步骤的专家共同努力，这样一个要求如此高和重要的项目才会成功。在我们的案例中，实现这一目标所需的技术诀窍在两个小组之间互补分配，其中一个小组位于奥地利维也纳(Monika Bradl和Hans Lassmann)，另一个小组位于德国慕尼黑(Klaus Dornmair)。因此，我们建议我们两个小组在DACH牵头机构行动下开展一个联合两国项目。这种联合项目具有特别重要的附加值，因为我们将能够直接比较我们从进展期多发性硬化症获得的数据和抗原与从复发缓解型多发性硬化症和极早期多发性硬化症样本中获得的数据和抗原(目前在DFG支持的项目中获得的数据，CRC TR128A5提案)。

Cells from the immune system (lymphocytes, for example T cells and B cells/plasma cells) are found in the brains of patients with multiple sclerosis (MS), located in areas where myelin sheaths are destroyed and where neurons degenerate. Moreover, these cells are found in MS brains at the very beginning of the disease, in the relapsing-remitting stage, and also in the progressive phase of the disease. Do these cells belong just to a few clones gone awry, or to many different ones?What do they recognize?Do immune cells found in the brain of very early MS recognize the same antigens as immune cells found later, in the progressive phase of the disease?Studies to find answers for these questions have not been done yet for patients with progressive MS where suitable material is extremely rare and often formaldehyde-fixed and paraffin-embedded (FFPE) deemed useless for further analysis. However, we have such precious samples, and we have the expertise to use FFPE tissue for molecular studies. Studies of pathogenic lymphocytes within the tissue require sophisticated techniques like next generation sequencing to learn more about their repertoire (which is, to learn how different their antibodies and T cell receptors are, and whether they belong to few or many different clones). Over the last years, also these techniques have been adjusted to FFPE samples. Disease-relevant cells are typically expanded within the tissue. Therefore, we will next clone and express the most abundant antibodies and T cell receptors identified in the progressive MS brains to find out what these molecules recognize. And finally, when we know this, we will go back to pathology to find out when and where this recognition occurs. Answers to these questions could help to better understand the driving forces of MS, and perhaps even to find better therapies for MS.It can be easily envisaged that such a demanding and important project will only be successful when specialists for all these steps jointly work together. In our case, the technical know-how needed to achieve this goal is complementarily distributed between two groups, on of them located in Vienna, Austria (Monika Bradl and Hans Lassmann), the other one located in Munich, Germany (Klaus Dornmair). Therefore, we propose a joint binational project of our two groups under the DACH lead agency action. Such a joint project has an especially important point of added value since we will be able to directly compare our data and antigens from progressive MS to those obtained from samples with relapsing remitting MS and with very early MS (data currently obtained in a project supported by DFG, proposal CRC TR 128 A5).