TCR Transduction for EBV Specific Immunotherapy

用于 EBV 特异性免疫治疗的 TCR 转导

• 批准号: 6607628
• 负责人: RIMAS J ORENTAS
• 金额: $ 23.63万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6607628
• 关键词: CD3 molecule; Epstein Barr virus; Hodgkin's disease; Retroviridae; T cell receptor; biological signal transduction; cytotoxic T lymphocyte; human tissue; molecular cloning; neoplasm /cancer immunology; neoplasm /cancer immunotherapy; receptor expression; recombinant proteins; transfection /expression vector; tumor antigens; virus antigen; virus related neoplasm /cancer

DESCRIPTION (provided by applicant): TCR TRANSDUCTION FOR EBV-SPECIFIC IMMUNOTHERAPY. Adoptive immunotherapy with polyclonal cytotoxic T cell lines (CTL) has met with clinical success in the treatment of post-transplant lymphoma, an Epstein-Barr virus (EBV)-associated malignancy that expresses the most immunodominant EBV latency antigens. This strategy is not applicable to two other EBV-associated malignancies, Hodgkin's disease (appx. 50 percent of cases are EBV-associated) and nasopharyngeal carcinoma (100 percent EBV-associated). These malignancies only express the EBV latency antigens LMP-1, LMP-2, and EBNA-1; none of which induce a strong immune response. These represent sub-dominant tumor-associated antigens. The goal of this project is to provide an immunotherapeutic option to patients suffering from these diseases by cloning individual T cell receptor molecules (TCR) that recognize LMP-1 and LMP-2 in an HLA-restricted manner, and introducing these recombinant TCR into HLA-A2 lymphocytes. Toward that goal, CTL clones specific for LMP-2 will be generated, the TCR alpha and beta chains molecularly cloned, and then transferred to retroviral expression vectors. These vectors will then be used to transduce CTL clones of known specificity as well as activated primary lymphocytes in bulk culture. The specific aims of this project seek to determine which TCRs are the best candidates for genetic transduction by comparing the CTL activity of the original cell to the lytic activity newly conferred upon the transduced cell. It remains to be determined whether it is the primary sequence of the TCR or the physiology of the transduced cell that determines the cytolytic activity conferred by the new receptor. We will also determine the structure of the TCR-CD3 complex in transduced cells, and in examining the bulk transduced lymphocyte population determine which cells are capable of expressing the transduced receptor. Should the retroviral vector used in these studies not give long-term expression of the transduced TCR, we also propose a newer generation of retroviral vectors that would be used instead. Once transduced, the newly expressed TCR-alpha and beta chains will have to compete with the endogenous TCR elements for association with the CD3 receptor complex and subsequent transit to the cell surface. Data obtained from this project will allow correlation between levels of retroviral gene transduction, mRNA expression, intracellular protein expression (the assembly of ICR subunits in the endoplasmic reticulum), cell surface expression of transduced TCR, and lytic function to be made. With a better understanding of these first principles of functional ICR assembly in primary lymphocytes, other malignancies with known tumor-associated antigens could be targeted by this approach as well.

描述（由申请人提供）：EBV特异性的TCR转导 免疫疗法。多克隆细胞毒性T细胞系的收养免疫疗法 （CTL）在移植后治疗方面取得了临床成功 淋巴瘤，一种爱泼斯坦 - 巴尔病毒（EBV）相关的恶性肿瘤，表达 大多数免疫主导的EBV潜伏期抗原。此策略不适用于 另外两个与EBV相关的恶性肿瘤，霍奇金氏病（Appx。50％ 病例与EBV相关）和鼻咽癌（100％ 与EBV相关）。这些恶性肿瘤仅表示EBV潜伏期抗原 LMP-1，LMP-2和EBNA-1；这些都没有引起强烈的免疫反应。这些 代表亚抑制肿瘤相关的抗原。这个项目的目标是 为患有这些的患者提供免疫治疗选择 通过克隆单个T细胞受体分子（TCR）来识别疾病 以HLA限制的方式LMP-1和LMP-2，并引入这些重组 TCR进入HLA-A2淋巴细胞。为此，针对LMP-2的CTL克隆 将产生，TCRα和β链分子克隆，然后 转移到逆转录病毒表达载体。然后将使用这些向量 传递已知特异性的CTL克隆和激活的主要克隆 大量培养中的淋巴细胞。该项目的具体目的是 确定哪些TCR是通过 将原始细胞的CTL活性与新的裂解活性进行比较 赋予转导的细胞。是否是 TCR的主要序列或转导细胞的生理学 确定新受体赋予的细胞溶解活性。我们也会 确定转导细胞中TCR-CD3复合物的结构，并在 检查散装转导的淋巴细胞种群确定哪些细胞是 能够表达转导的受体。应该逆转录病毒载体 在这些研究中使用的不长期表达转导的TCR，我们 还提出了一种新一代的逆转录病毒载体 反而。一旦转导，新表达的TCR-alpha和beta链将 必须与内源性TCR元素竞争与CD3相关 受体复合物，随后转移到细胞表面。从中获得的数据 该项目将允许逆转录病毒基因之间的相关性 转导，mRNA表达，细胞内蛋白表达（组装 内质网中的ICR亚基），细胞表面表达 转导的TCR和要产生的裂解功能。更好地理解 这些功能性ICR组装的第一原理在原发性淋巴细胞中，其他 具有已知肿瘤相关抗原的恶性肿瘤可能是针对的 方法也是如此。