T cell receptor gene vectors for EBV disease

描述（由申请人提供）：本申请的目的是确定表达特异性针对EBV潜伏抗原LMP-2的HLA-A2或HLA-23，24限制性T细胞受体（TCR）的慢病毒工程化T细胞是否能够识别由EBV转化的原代B淋巴细胞（B-LCL）表达的该抗原以产生细胞溶解效应。在产生和评估这些载体时，我们将产生一组独特的用于EBV相关疾病的基因治疗载体，确定哪些外周血亚群用作转导的最佳靶标，并且还确定转导的T细胞是否与未转导的T细胞经受相同的免疫对照。记录慢病毒转导的T细胞的正常免疫生物学将增强已经从这些载体在HIV患者的I期试验中的临床使用中获得的信心。医学上重要的并且被认为适合免疫控制的爱泼斯坦巴尔病毒（EBV）相关疾病包括AIDS相关淋巴瘤、霍奇金淋巴瘤、移植后淋巴瘤和鼻咽癌。这些EBV相关恶性肿瘤中的每一种都表达EBV编码的潜伏膜蛋白-2（LMP-2）。Lentigen的合作者Rimas Orentas博士帮助开创了使用过继免疫疗法治疗EBV相关恶性肿瘤的先河。在目标1中，我们将开发表达由Orentas实验室克隆的TCR的自失活（SIN）慢病毒载体，其靶向LMP-2的HLA-A2限制性（残基131-139）或HLA-23，24限制性（残基425-433）表位。在以前的工作中，Orentas实验室证明了这些TCR能够使用逆转录病毒载体在原代人类T细胞中功能性表达。重要的是，这是证明裂解B-LCL（与肽脉冲靶或经工程改造以表达LMP-2的靶相对的具有高生物学相关性的靶细胞）的能力的第一份报告。然而，逆转录病毒载体是次优的，即使在实验室规模上也难以生产高滴度储备液。利用Lentigen的研究和开发能力，我们建议使用最先进的慢病毒系统生成新的载体，这将使临床和研究界掌握治疗EBV相关恶性肿瘤的有效工具。在过去的几年里，使用TCR治疗疾病已经从一个推测性的研究课题发展到具体的临床方案。我们建议用我们克隆的LMP-2特异性TCR做同样的事情。我们的最终目标是为患有EBV相关恶性肿瘤的患者提供免疫选择。

DESCRIPTION (provided by applicant): The goal of this application is to determine if lentiviral-engineered T cells, that express either an HLA-A2 or an HLA-23,24 restricted T cell receptor (TCR) specific for the EBV latency antigen LMP-2, are capable of recognizing this antigen as expressed by EBV transformed primary B lymphocytes (B-LCL) to a cytolytic effect. In producing and evaluating these vectors we will generate a unique set of gene therapy vectors for EBV- associated disease, determine which peripheral blood subsets serve as the optimal target for transduction and also determine whether transduced T cells are subject to the same immune controls as non-transduced T cells. Documenting the normal immunobiology of lentiviral transduced T cells will enhance the confidence already being gained from clinical use of these vectors in phase I trials for HIV patients. Epstein Barr virus (EBV)-associated diseases that are medically significant and thought to be amenable to immune control include AIDS-associated lymphoma, Hodgkin lymphoma, post-transplant lymphoma, and nasopharyngeal carcinoma. Each of these EBV-associated malignancies expresses the EBV-encoded latency membrane protein-2 (LMP-2). Lentigen's collaborator, Dr. Rimas Orentas, has helped pioneer the used of adoptive immunotherapy to treat EBV-associated malignancy. In Aim 1 we will develop self-inactivating (SIN) lentiviral vectors that express TCRs cloned by the Orentas lab, that target an HLA-A2-restricted (residues 131-139) or HLA-23,24-restricted (residues 425-433) epitope of LMP-2. In previous work the Orentas lab demonstrated the ability of these TCRs to be functionally expressed in primary human T cells using a retroviral vector. Importantly, this was the first report to demonstrate the ability to lyse B-LCL (a target cell of high biological relevance as opposed to peptide- pulsed targets or targets engineered to express LMP-2). However, the retroviral vector was suboptimal and production of high-titer stock difficult on even a laboratory scale. Using the research and development capability of Lentigen, we propose here to generate new vectors, using a state-of-the-art lentiviral system, that will place in the hands of the clinical and research community effective tools to treat EBV-associated malignancy. Over the last few years the use of TCRs to treat disease has gone from a speculative research topic to concrete clinical protocols. We propose to do the same with our cloned LMP-2 specific TCRs. Our ultimate goal is to provide an immunotherapeutic options to patients suffering from EBV-associated malignancy.