Molecular and cellular mechanisms of synapse-mediated spread of Epstein Barr virus: overcoming the CD21-restricted cellular tropism.

Epstein Barr 病毒突触介导传播的分子和细胞机制：克服 CD21 限制的细胞向性。

• 批准号: MR/J002046/1
• 负责人: Claire Shannon-Lowe
• 金额: $ 56.45万
• 依托单位: University of Birmingham
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2012
• 资助国家: 英国
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FJ002046%2F1
• 关键词: Molecular; cellular; mechanisms; synapse; mediated

Epstein Barr Virus (EBV) is a common, orally-transmitted virus, infecting most of the human population. EBV maintains a silent infection for the lifetime of the host by hiding in a subset of lymphocytes called B cells. EBV infection of B cells in the laboratory has been extensively studied. In the laboratory, infected B cells are efficiently transformed into continuously growing cells with the potential to cause cancer. This happens only rarely in the body, but when it does can cause B cell cancers including Hodgkin lymphoma. One reason why B cell cancers are so rare is because infection is kept under control by immune responses involving other cells, including T and natural killer (NK) cells.The mechanisms of EBV entry into B cells have been extensively studied. Proteins on the virus (gp350 and gp42) interact with virus receptors on the cell surface (CD21, HLA class II respectively) to bind the EBV to the B cell. Thereafter further virus proteins (gp82, gp25) trigger virus internalisation. However, infection of other cell types, including epithelial, T and NK cells that do not express the CD21 are also observed. These infections are associated with either virus transmission to a new susceptible host (epithelial), or are associated with diseases and malignancies of epithelial, T and NK cells.Epithelial cells line the oral cavity (oropharynx) and create a barrier for entry of EBV into an uninfected host, and for escape of EBV from an infected host for transmission to a new susceptible host. Epithelial cells are thought to be the major site of EBV replication in the oropharynx; however they do not express CD21, and EBV infection of epithelial cells in the laboratory is poor. I have previously shown that infection of epithelial cells can be greatly enhanced by first binding the virus to B cells. EBV binding to CD21 triggers adhesion molecules and EBV on the B cell surface to polarise then 'adhere' to the epithelial cells, bringing EBV into close contact with epithelial cells and enabling efficient infection. This process is probably similar to the situation in the oropharynx, where epithelial cells and lymphocytes, including B cells, are in close contact with each other.In our model of EBV infection of epithelial cells, we suggest EBV on the B cells may come from fully matured B cells, called plasma cells, which when infected with EBV produce new infectious virus. This EBV may be released from the plasma cell to interact with the resting B cell as above. Or, as increasingly observed with other viruses that infect lymphocytes (like HIV), the plasma cell may interact directly with the epithelial cell, cause EBV inside the plasma cell to relocate to the point of cell-to-cell contact and transmit directly from the plasma cell to the epithelial cell, thereby evading anti-EBV antibodies. Here, I intend to extend my studies to understand how EBV can infect epithelial cells using plasma cells as the virus donor, and determine the molecular requirements of epithelial cells and EBV for efficient infection, so we can understand how to disrupt EBV transmission with vaccines.EBV associated T and NK cell diseases range from illnesses that appear to be a consequence of an over-active immune system (EBV-associated haemophagocytic lymphohistiocytosis, chronic active EBV), to aggressive malignancies (extranodal T/NK cell lymphoma, aggressive NK cell leukaemia).T and NK cells do not express CD21 and we do not understand how these cells can be infected with EBV. Using two different methods, I aim to determine how infection of these cells is achieved: (1) Using plasma cell interaction with T and NK cells, and (2) infecting CD21-positive immature T and NK cells. With this new information, we will be able to develop ideas of how the virus may be causing the T and NK cell diseases, and begin develop new treatments for these treatment-resistant diseases.

爱泼斯坦-巴尔病毒(EBV)是一种常见的口腔传播病毒，感染大多数人类人群。EBV通过隐藏在称为B细胞的淋巴细胞亚群中，在宿主的一生中保持沉默的感染。EB病毒在实验室对B细胞的感染已有广泛研究。在实验室中，被感染的B细胞被有效地转化为具有致癌潜力的持续生长的细胞。这种情况在人体内很少发生，但一旦发生，就会导致B细胞癌，包括霍奇金淋巴瘤。B细胞癌如此罕见的一个原因是感染受到包括T细胞和自然杀伤(NK)细胞在内的其他细胞的免疫反应的控制。EBV进入B细胞的机制已经得到了广泛的研究。病毒上的蛋白(gp350和gp42)与细胞表面的病毒受体(分别为CD21和HLAII类)相互作用，将EBV与B细胞结合。此后，更多的病毒蛋白(gp82、gp25)触发病毒内化。然而，也可以观察到其他细胞类型的感染，包括不表达CD21的上皮细胞、T细胞和NK细胞。这些感染与病毒传播到新的易感宿主(上皮)有关，或者与上皮细胞、T细胞和NK细胞的疾病和恶性肿瘤有关。上皮细胞排列在口腔(口咽部)并为EBV进入未感染的宿主和EBV从感染的宿主逃逸而传播到新的敏感宿主创造屏障。上皮细胞被认为是EBV在口咽部复制的主要部位，但它们不表达CD21，实验室中EBV对上皮细胞的感染很差。我之前已经证明，通过首先将病毒与B细胞结合，可以极大地增强上皮细胞的感染。EBV与CD21结合后，会触发B细胞表面的黏附分子和EBV极化，然后与上皮细胞黏附，使EBV与上皮细胞密切接触，从而实现有效的感染。这一过程可能类似于口咽部的情况，在口咽上皮细胞和淋巴细胞，包括B细胞，密切接触。在我们的上皮细胞EBV感染模型中，我们认为B细胞上的EBV可能来自完全成熟的B细胞，称为浆细胞，当EBV感染时，它会产生新的传染性病毒。如上所述，这种EBV可以从浆细胞中释放出来，与静止的B细胞相互作用。或者，正如越来越多地观察到其他感染淋巴细胞的病毒(如艾滋病毒)，浆细胞可能直接与上皮细胞相互作用，导致浆细胞内的EBV重新定位到细胞与细胞的接触点，并直接从浆细胞传播到上皮细胞，从而躲避抗EBV抗体。在这里，我打算扩大我的研究范围，以了解EBV如何使用浆细胞作为病毒供体感染上皮细胞，并确定上皮细胞和EBV有效感染的分子要求，以便我们能够了解如何通过疫苗阻断EBV的传播。EBV相关的T和NK细胞疾病从似乎是免疫系统过度活跃的疾病(EBV相关的吞噬血细胞淋巴组织细胞增生症，慢性活动性EBV)到侵袭性恶性肿瘤(结外T/NK细胞淋巴瘤，侵袭性NK细胞白血病)。T和NK细胞不表达CD21，我们不了解这些细胞如何感染EBV。通过两种不同的方法，我的目的是确定这些细胞是如何感染的：(1)通过浆细胞与T细胞和NK细胞相互作用，以及(2)感染CD21阳性的未成熟T细胞和NK细胞。有了这些新的信息，我们将能够提出病毒可能如何导致T和NK细胞疾病的想法，并开始为这些耐药疾病开发新的治疗方法。

项目成果

Epstein Barr virus entry; kissing and conjugation.

• DOI: 10.1016/j.coviro.2013.12.001
• 发表时间: 2014-02
• 期刊: Current opinion in virology
• 影响因子: 5.9
• 作者: C. Shannon-Lowe;M. Rowe

Cooperation between Epstein-Barr virus immune evasion proteins spreads protection from CD8+ T cell recognition across all three phases of the lytic cycle.

• DOI: 10.1371/journal.ppat.1004322
• 发表时间: 2014-08
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Quinn LL;Zuo J;Abbott RJ;Shannon-Lowe C;Tierney RJ;Hislop AD;Rowe M
• 通讯作者: Rowe M

Early T Cell Recognition of B Cells following Epstein-Barr Virus Infection: Identifying Potential Targets for Prophylactic Vaccination.

• DOI: 10.1371/journal.ppat.1005549
• 发表时间: 2016-04
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Brooks JM;Long HM;Tierney RJ;Shannon-Lowe C;Leese AM;Fitzpatrick M;Taylor GS;Rickinson AB
• 通讯作者: Rickinson AB

Counteracting effects of cellular Notch and Epstein-Barr virus EBNA2: implications for stromal effects on virus-host interactions.

• DOI: 10.1128/jvi.01431-14
• 发表时间: 2014-10
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Rowe M;Raithatha S;Shannon-Lowe C
• 通讯作者: Shannon-Lowe C

Unexpected patterns of Epstein-Barr virus transcription revealed by a high throughput PCR array for absolute quantification of viral mRNA.

• DOI: 10.1016/j.virol.2014.10.030
• 发表时间: 2015-01-01
• 期刊: VIROLOGY
• 影响因子: 3.7
• 作者: Tierney, Rosemary J.;Shannon-Lowe, Claire D.;Fitzsimmons, Leah;Bell, Andrew I.;Rowe, Martin
• 通讯作者: Rowe, Martin