CLINICAL TRIAL: ADMINISTRATION OF PERIPHERAL BLOOD T-CELLS AND EBV SPECIFIC CTLS

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。 首要目标 目的：评价CD45单抗去除淋巴组织后，递增剂量14g2a.zeta嵌合受体转导的自体EBV特异性细胞毒性T淋巴细胞(EBV-CTL)和14g2a.zeta转导的自体外周血T细胞的安全性。 次要目标 为了确定这两种输注细胞类型在体内的存活和功能的差异，特别是确定嵌合受体转导的EBV-CTL是否比转导的外周血T细胞存活时间更长。 检测转导的外周血T细胞和EBV特异性CTL的体内抗肿瘤作用。 利用免疫系统来根除疾病的概念已经有了 神经母细胞瘤研究的长期货币，得到了 肿瘤的生物学行为，包括年幼儿童的自发消退。此外，由于神经母细胞瘤来源于胚胎神经外胚层，它表达在非胚胎组织中不广泛检测到的抗原11，并可能过度表达其他细胞抗原。最后，在动物和人类中，神经母细胞瘤细胞在体外和体内都对细胞毒效应机制敏感。13-15神经母细胞瘤的临床免疫治疗有多种形式，大致可分为疫苗/细胞因子方法或治疗性单抗的使用。 通过测量外周血液中转基因的水平，我们将能够估计转基因T细胞存活的总体动力学，并确定在体内是否发生了扩增或持续。即使CTL来源的信号确实存在，我们检测到的转基因阳性细胞也可能失去对EBV和神经母细胞瘤的一种或另一种特异性。我们将能够使用FACS对四聚体阳性细胞进行分选，以发现它们是否保持了对抗恶性和EBV感染的靶细胞的双功能活性。

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. Primary Objective To evaluate the safety of escalating doses of 14g2a.zeta chimeric receptor transduced autologous EBV specific cytotoxic T-lymphocytes (EBV-CTL)and 14g2a.zeta transduced autologous peripheral blood T-cells administered to patients with Neuroblastoma who have been lymphodepleted by CD45 monoclonal antibodies (MAbs). Secondary Objectives To determine the differential survival and function of these two infused cell-types in vivo, in particular to determine if chimeric receptor transduced EBV-CTLs survive longer than transduced peripheral-blood T-cells. To determine anti-tumor effects of transduced peripheral blood T-cells and EBV specific CTLs in vivo. The concept of exploiting the immune system to eradicate disease has had a long currency in neuroblastoma research, supported by many aspects of the tumor's biologic behavior, including spontaneous regression in younger children. In addition, because neuroblastoma is derived from embryonic neuroectoderm, it expresses antigens not widely detected in non-embryonic tissues11, and may overexpress other cellular antigens. Finally, in animals and in man, neuroblastoma cells are susceptible to cytotoxic effector mechanisms both in vitro and in vivo.13-15 Clinical immunotherapy for neuroblastoma has taken many forms which may be loosely classified into vaccine / cytokine approaches or the use of therapeutic monoclonal antibodies. By measuring the level of the transgene in peripheral blood we will be able to estimate the overall kinetics of gene- modified T-cell survival and determine whether expansion or persistence occures in vivo. Even if CTL derived signal does persist it is possible that the transgene-positive cells we detect will have lost one or other of their specificities for EBV and neuroblastoma. We will be able to use FACS sorting of tetramer positive cells to discover whether or not they retain their bifunctional activity against malignant and EBV-infected target cells.