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EBV潜伏期抗原LMP-2特异性HLA-A2或hla - 232,24限制性T细胞受体（TCR）的慢病毒工程T细胞是否能够识别EBV转化的原代B淋巴细胞（B- lcl）表达的这种抗原，从而产生细胞溶解作用。在生产和评估这些载体的过程中，我们将为EBV相关疾病生成一套独特的基因治疗载体，确定哪些外周血亚群作为转导的最佳靶标，并确定转导的T细胞是否受到与非转导T细胞相同的免疫控制。记录慢病毒转导T细胞的正常免疫生物学将增强这些载体在HIV患者的I期临床试验中已经获得的信心。eb病毒（EBV）相关疾病在医学上具有重要意义，被认为可以免疫控制，包括艾滋病相关淋巴瘤、霍奇金淋巴瘤、移植后淋巴瘤和鼻咽癌。这些ebv相关的恶性肿瘤都表达ebv编码的潜伏期膜蛋白-2 （LMP-2）。Lentigen的合作者Rimas Orentas博士率先使用过继免疫疗法治疗ebv相关的恶性肿瘤。在Aim 1中，我们将开发表达由Orentas实验室克隆的tcr的自灭活（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。在之前的工作中，奥伦塔斯实验室使用逆转录病毒载体证明了这些tcr在原代人T细胞中功能性表达的能力。重要的是，这是第一个证明能够裂解B-LCL（与肽脉冲靶细胞或工程靶细胞表达LMP-2相反，具有高生物学相关性的靶细胞）的报告。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的存量。利用Lentigen的研发能力，我们建议使用最先进的慢病毒系统生成新的载体，这将为临床和研究界提供有效的工具来治疗ebv相关的恶性肿瘤。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变成了具体的临床方案。我们建议对克隆的LMP-2特异性tcr进行同样的处理。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治疗选择。

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.