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的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 这是一个采购专用协议 这是以下GCRC支持的研究的配套方案： H-9935、6676、9454、8216使用从合格受试者获得的血液，将能够产生EBV特异性细胞毒性T淋巴细胞（CTL）。 本方案的目的是获得用于制备EBV特异性细胞毒性T淋巴细胞（CTL）的组织（血液）。如果成功的话，这些细胞将来可能会被送回给患者，如果他们有资格参加正在进行的EBV-CTL研究。 EB病毒（Epstein-Barr virus，EBV）是一种能够感染人B淋巴细胞并使其永生化的人嗜淋巴细胞疱疹病毒。在通过唾液交换发生的原发性感染中，EBV感染口咽上皮细胞，口咽上皮细胞支持裂解性复制并允许病毒传播到循环B细胞。一旦进入B细胞，EBV基因组就保持潜伏形式。体外感染EBV的B细胞经历 转化是潜伏相关转化蛋白表达的结果。这些B细胞总是表达许多EBV基因产物，包括核抗原EBNA 1、EBNA 2、EBNA 3A、EBNA 3 B、EBNA 3C、EBNA LP和潜伏膜蛋白LMP 1和LMP 2。表达这些潜伏相关转化蛋白的EBV感染细胞的增殖受特异性T细胞控制。因此，在大多数人群中，原发性感染无症状或导致轻度和自限性疾病（传染性单核细胞增多症，IM）。感染后，个体是终身病毒携带者。与具有正常免疫系统的患者不同，先天性或获得性免疫缺陷患者非常容易发生EBV驱动的淋巴组织增生性疾病（EBV-LPD），因为EBV的再活化不太受细胞介导的反应的严格控制。有证据表明，EBV特异性CD 8 + CTL是对抗EBV感染的B细胞生长的最重要的防御机制。这些细胞识别来自与B细胞表面上的MHC分子结合表达的病毒抗原的肽片段。免疫抑制个体中T细胞活性降低或抑制的状态干扰免疫监视系统并增加EBV驱动的感染细胞增殖的风险，从而允许具有生长优势的B细胞克隆出现并导致一个或多个克隆群体。与EBV的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，其中患者表现出慢性疲劳、发热、肝脾肿大和淋巴结病。患者的特征是对病毒衣壳抗原的抗体滴度升高，对EBNA的抗体低或缺乏，血清或其他体液中有游离病毒。EBV也与其他恶性肿瘤有关，并且在40%的霍奇金病患者和大多数鼻咽癌病例的恶性细胞中发现。在动物和人中的研究已经显示，EBV特异性T细胞可以离体产生以产生抗EBV感染的细胞免疫应答。我们已经在骨髓移植后的患者中使用了这种方法，其中供体来源的EBV特异性CTL在预防和治疗EBV淋巴细胞增殖方面都是有效的。我们现正将此方法扩展至其他EB病毒相关疾病，包括实体器官移植后出现的EB病毒淋巴瘤、EB病毒阳性何杰金氏病、鼻咽癌及严重慢性EB病毒感染。

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.