PROCUREMENT OF TISSUE FOR MAKING EPSTEIN-BARR VIRUS (EBV) SPECIFIC CYTOTOXIC T

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 这是一项仅限采购的协议 这是以下GCRC支持的研究研究的配套方案： H-9935、6676、9454、8216使用合格受试者的血液，将能够产生EBV特异性细胞毒性T淋巴细胞(CTL)。 该方案的目的是获取组织(血液)，用于制造EBV特异性细胞毒性T淋巴细胞(CTL)。如果成功，这些细胞将来可能会被回馈给患者，如果他们有资格参加正在进行的EBV-CTL研究之一。 EB病毒(Epstein-Barr Virus，EBV)是一种人类嗜淋巴疱疹病毒，能感染并永生化人类B淋巴细胞。在原发感染中，EBV通过唾液交换发生，感染支持裂解复制的口咽上皮细胞，并允许病毒传播到循环中的B细胞。一旦进入B细胞，EBV基因组就保持潜伏状态。在体外感染EBV的B细胞经历 转化是潜伏期相关转化蛋白表达的结果。这些B细胞总是表达多种EBV基因产物，包括核抗原EBNA1、EBNA2、EBNA3A、EBNA3B、EBNA 3C、EBNA LP以及潜伏膜蛋白LMP1和LMP2。表达这些潜伏期相关转化蛋白的EBV感染细胞的增殖受特定T细胞的控制。因此，在大多数人群中，原发感染是无症状的或导致轻微的自限性疾病(传染性单核细胞增多症，IM)。感染后，个人是终生病毒携带者。与免疫系统正常的患者不同，先天性或获得性免疫缺陷的人非常容易患上EBV驱动的淋巴增殖性疾病(EBV-LPD)，因为EBV的重新激活不太受细胞介导的反应的严格控制。有证据表明，EBV特异性CD8+CTL是抵抗EBV感染B细胞生长的最重要的防御机制。这些细胞识别来自病毒抗原的多肽片段，这些片段与B细胞表面的MHC分子相关联。免疫抑制个体中T细胞活性降低或抑制的状态会干扰免疫监视系统，增加受感染细胞由EBV驱动的增殖风险，使具有生长优势的B细胞克隆出现，并导致一个或多个克隆群体。与EBV免疫控制缺陷相关的另一种综合征是严重的慢性活动性EBV(SCAEBV)感染综合征，患者表现为慢性疲劳、发热、肝脾肿大和淋巴结病。患者的特点是对病毒衣壳抗原的抗体效价升高，对EBNA和血清或其他体液中的游离病毒的抗体很低或不存在。EBV还与其他恶性肿瘤有关，在40%的霍奇金病患者和大多数鼻咽癌患者的恶性细胞中发现EBV。在动物和人类身上的研究表明，EBV特异性T细胞可以在体外产生抗EBV感染的细胞免疫反应。我们在骨髓移植后的患者中使用了这种方法，在这些患者中，供体来源的EBV特异性CTL在预防和治疗EBV淋巴增殖方面都是有效的。我们现正将此方法扩展至其他与EBV有关的疾病，包括实体器官移植后出现的EBV淋巴瘤、EBV阳性霍奇金病、鼻咽癌和严重的慢性EBV感染。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This is a procurement only protocol This is a companion protocol to the following GCRC supported research studies: H-9935, 6676, 9454, 8216 Using blood obtained from eligible subjects, will enable the ability to produce EBV specific cytotoxic T lymphocytes (CTLs). The objective of this protocol is to obtain tissue (blood) for the purpose of making EBV specific cytotoxic T lymphocytes (CTLs). If successful, these cells may be given back to the patient in the future should they become eligible for one of the EBV-CTL research studies being conducted. Epstein-Barr virus (EBV) is a human lymphotropic herpes virus able to infect and immortalize human B lymphocytes. In the primary infection, occurring via saliva exchange, EBV infects oropharyngeal epithelial cells that support the lytic replication and allow the transmission of the virus to circulating B cells. Once in the B cells, the EBV genome is maintained in a latent form. B cells infected with EBV in vitro undergo transformation as result of the expression of latency-associated transforming proteins. These B cells always express a number of EBV gene products including the nuclear antigens EBNA1, EBNA2, EBNA3A, EBNA3B, EBNA 3C, EBNA LP and the latent membrane proteins LMP1 and LMP2. The proliferation of EBV-infected cells expressing these latency-associated transforming proteins is controlled by specific T cells. Hence, in a majority of the population, primary infection is asymptomatic or results in a mild and self-limiting illness (infectious mononucleosis, IM). Following infection, individuals are life-long virus carriers. Unlike patients with a normal immune system, people with congenital or acquired immunodeficiency are highly susceptible to the development of EBV driven lymphoproliferative disease (EBV-LPD) since the reactivation of EBV is less tightly controlled by a cell-mediated response. Evidence suggests that EBV specific CD8+ CTLs are the most important defense mechanism against outgrowth of EBV infected B cells. These cells recognize peptide fragments derived from viral antigens expressed in association with MHC molecules on the surface of B cells. The state of reduced or suppressed T cell activity in immunosuppressed individuals interferes with the immunosurveillance system and increases the risk of EBV-driven proliferation of the infected cells, allowing Bcell clones with a growth advantage to emerge and result in one or more clonal populations. Another syndrome associated with defective immune control of EBV is severe chronic active EBV (SCAEBV) infection syndrome where patients present with chronic fatigue, fever, hepatosplenomegaly and lymphoadenopathy . Patients characteristically have elevated antibody titers to the viruscapsid antigen and low or absent antibody to EBNA and free virus in serum or other body fluids. EBV is also associated with other malignancies and is found in the malignant cells of 40% of patients with Hodgkin Disease, and most cases of nasopharyngeal carcinoma. Studies in animals and humans have shown that EBV specific T-cells can be generated ex-vivo to produce an anti- EBV infected cell immune response. We have used this approach in patients post bone marrow transplant where donor derived EBV-specific CTLs have been effective both in preventing and treating EBV lymphoproliferation. We are now extending this approach to other EBV-associated diseases including EBV lymphomas arising after solid organ transplant, EBV positive Hodgkin disease, nasopharyngeal carcinoma and severe chronic EBV infection.