UNDERSTANDING THE ROLE OF EPSTEIN BARR VIRUS IN T CELL AND NATURAL KILLER CELL LYMPHOPROLIFERATIONS AND MALIGNANCIES

爱泼斯坦巴尔病毒（EBV）是一种常见的疱疹病毒，在每个人的一生中都会感染大多数人。绝大多数人都不知道他们已经被感染了，因为病毒被他们的免疫系统严密控制着。虽然EBV以引起腺热最为人所知，但它也与少数个体的癌症有关。这些癌症通常发生在被感染的细胞中，作为病毒自然生命周期的一部分，B淋巴细胞和上皮细胞。然而，在极少数情况下，EBV也可以感染不参与病毒生命周期的细胞，如T淋巴细胞和自然杀伤（NK）细胞。这些细胞类型的感染总是导致严重的、危及生命的疾病，包括癌症。这些疾病极难治疗，临床结果很差，通常以生存数周到数月来衡量。迫切需要更好地了解这些ebv相关T/NK细胞疾病的病理，以便开发新的有效的个性化治疗方法。由于这些疾病的罕见性和我们无法在实验室感染T/NK细胞，了解这些疾病的进展非常缓慢。然而，我们已经解决了这两个问题。我们目前有一个英国ebv相关T/NK细胞疾病患者的血液样本库准备进行分析。此外，我们还优化了培养条件，并从患者的样本中产生了匹配的ebv阳性T/NK细胞系。最后，也许也是最重要的一点，我们已经成功地在实验室中复制感染T细胞，这将使我们第一次能够研究EBV如何触发这些T/NK细胞疾病并诱导其转化为恶性肿瘤。我们有四个主要目标。(1)利用患者血液，我们从微环境中鉴定出白细胞和化学信号（细胞因子），这是帮助患者感染ebv的T/NK细胞在培养中生长所必需的。我们计划在实验室中复制这些条件，以帮助新感染的T/NK细胞生长，并确定EBV，周围细胞和细胞因子如何促进新感染的T/NK细胞的生长。(2)一旦生长条件优化，我们将研究特异性EBV基因产物如何能够刺激EBV相关T/NK细胞疾病中观察到的慢性炎症。(3)通过在细胞培养中复制EBV感染和慢性炎症，我们将研究EBV和慢性炎症如何驱动感染细胞无限生长并失调抗EBV免疫。(4)建立了一种新的分析工具，用EBV标记物和细胞类型标记物标记感染的细胞。我们可以扩大标记物的范围，以识别ebv感染细胞上潜在的新药靶点，包括治疗性抗体，如抗ccr4 (mogamulizumab)，目前正处于治疗外周T细胞淋巴瘤的2期临床试验中。此外，如上所述，我们可以在药物存在的情况下培养匹配的ebv感染的离体T/NK细胞，以确定对药物的敏感性。这是同类研究中首次通过结合离体患者样本和原发感染的T/NK细胞来分析EBV对EBV相关T/NK细胞疾病的贡献。我们预计，通过了解EBV如何直接或与周围微环境的其他细胞和细胞因子联合引起疾病，并通过监测体外和体内的疾病进展，我们将更好地了解替代的新治疗方案。最后，基于这项研究的发现，我们建立的检测潜在新疗法的方法将对ebv相关T/NK细胞疾病患者群体有更广泛的益处，如果这项研究进入试验阶段。研究结果将确定个性化治疗方案的潜在药物。

Epstein Barr virus (EBV) is a common herpesvirus that infects the majority of the human population for the lifetime of each individual. The vast majority of individuals are unaware that they have been infected because the virus is kept under tight control by their immune system. Although EBV is most well known for causing glandular fever, it is also associated with cancers in a small number of individuals. These cancers usually occur in the cells infected as part of the natural life cycle of the virus, B lymphocytes and epithelial cells. However, on rare occasions EBV can also infect cells that are not involved in the lifecycle of the virus, T lymphocytes and Natural killer (NK) cells. Infection of these cell types always results in serious, life-threatening diseases including cancers. These diseases are extremely difficult to treat and have a very poor clinical outcome, usually measured in survival from weeks to months. There is an urgent need to better understand the pathology of these EBV-associated T/NK cell diseases in order to develop new effective, personalized therapies.Progress into understanding these diseases has been extremely slow, due to the rarity of the diseases and our inability to infect T/NK cells in the lab. However, we have addressed both these issues. We currently have a bank of blood samples from UK patients with EBV-associated T/NK cell diseases ready for analysis. Furthermore we have optimized the culture conditions and have generated matching EBV-positive T/NK cell lines from the patients' samples. Finally, and perhaps most importantly, we have managed to reproducibly infect T cells in our laboratory, which for the first time will allow us to examine how EBV triggers these T/NK cell diseases and induces their transformation into malignancy. We have four key objectives. (1) Using patient blood, we have identified white blood cells and chemical signals (cytokines) from the microenvironment which are required to help the patients' EBV-infected T/NK cells to grow in culture. We plan to replicate these conditions in the lab to help the newly-infected T/NK cells grow and determine how EBV, the surrounding cells and cytokines contribute to the growth of newly-infected T/NK cells. (2) Once the growth conditions are optimized, we will examine how specific EBV gene products able to stimulate the chronic inflammation observed in the EBV-associated T/NK cell diseases. (3) By replicating the EBV infection and chronic inflammation in our cell cultures, we will examine how EBV and chronic inflammation drive the infected cells to grow indefinitely and dysregulate the anti-EBV immunity. (4) We have established a new analysis tool, which labels the EBV-infected cells with markers for EBV and markers for the cell type. We can expand the range of markers to identify potential new drug targets on the EBV-infected cell to include therapeutic antibodies such as anti-CCR4 (mogamulizumab) currently in phase 2 clinical trial for peripheral T cell lymphoma. Furthermore, we can culture the matching ex-vivo EBV-infected T/NK cells, as above, in the presence of the drug to determine sensitivity to the drug. This is the first study of its kind to analyse the contribution of EBV to the EBV-associated T/NK cell diseases by combining ex-vivo patient samples and primary infected T/NK cells. We anticipate that by understanding how EBV causes the diseases, either directly or in combination with other cells and cytokines of the surrounding microenvironment, and by monitoring disease progression both in vitro and in vivo, we will develop a better understanding of alternative novel therapeutic options. Finally, the assays we have established to examine the potential new treatments will have wider benefit to the EBV-associated T/NK cell disease patient groups if the study were taken to trial, based on the findings of this study. The results will identify potential drugs for personalized treatment options.