EBV Specific T-cells from 3rd party donors for treatment of EBV-associated malign

描述（由申请人提供）：eb病毒诱导的淋巴细胞增生性疾病和EBV+淋巴瘤是同种异体造血细胞（HSCT）和器官移植（SOT）接受者以及严重遗传和获得性免疫缺陷患者发病率和死亡率的重要原因。EBV+霍奇金病（HD）、EBV+噬血细胞淋巴组织细胞增多症、EBV+鼻咽癌和EBV+平滑肌肉瘤也是一类目前化疗往往无效的恶性肿瘤。体外生成的供体来源的EBV特异性t细胞的过继治疗在治疗和预防EBV淋巴瘤合并异体造血干细胞移植方面是有效的。然而，由于缺乏生产t细胞所需的专门设施，以及需要时间（约65天）来产生足够数量、ebv特异性和缺乏同种异体反应性的ebv特异性t细胞，应用受到限制。利用EBV特异性t细胞进行有效治疗也可能受到以下因素的阻碍或阻碍：供体细胞用于t细胞繁殖的可用性有限，供体体内缺乏对病毒的预先敏化，或供体t细胞在肿瘤表达HLA等位基因的情况下无法识别EBV。根据我们的初步研究，我们假设从预先建立的t细胞系库中适当选择第三方供体获得的ebv特异性t细胞可以解决这些限制。因此，在目标1中，我们将：开展一项II期试验，在匹配2个HLA等位基因和适当的HLA限制的基础上，选择来自第三方供体的EBV特异性t细胞，用于治疗EBV淋巴瘤合并异体造血干细胞移植或器官移植或其他EBV相关恶性肿瘤伴免疫缺陷的患者。在这项试验中，我们将把临床反应和观察到的EBV DNA循环水平的变化与EBV特异性t细胞在过继转移后的体内生长、扩增和存活的定量评估联系起来。在目标2中，我们将建立一个大型的，遗传多样性的ebv特异性t细胞系库，特别同意第三方使用。300个t细胞系中的每一个都是在GMP条件下生产的，这些t细胞系是由同意的捐赠者产生的，并且是该银行的候选细胞系，其特点是其HLA基因型，EBV特异性，缺乏同种异体反应性和必要的微生物无菌性。我们计划确定每个品系中EBV特异性t细胞的HLA限制，以便允许为特定患者选择受患者肿瘤共有HLA等位基因限制的EBV t细胞。我们还将鉴定含有LMP-1和/或LMP-2特异性t细胞的细胞系，这些细胞系可用于治疗选择性表达这些抗原的EBV+恶性肿瘤。我们期望这项II期试验将提供启动III期研究所需的数据，将该疗法与化疗和/或Rituxan进行比较。具有良好特征的ebv特异性t细胞库也应该为此类试验提供广泛适用和可获得的资源。

DESCRIPTION (provided by applicant): Epstein-Barr virus induced lymphoproliferative disease and EBV+ lymphomas are a significant cause of morbidity and mortality in recipients of allogenic hematopoietic cell (HSCT) and organ transplants (SOT) as well as patients with severe genetic and acquired immunodeficiencies. EBV+ Hodgkin's disease (HD), EBV+ hemophagocytic lymphohistiocytosis, EBV+ nasopharyngeal carcinoma and EBV+ leiomyosarcomas also constitute a group of malignancies for which current chemotherapy is often ineffective. Adoptive therapy with donor-derived EBV-specific T-cells generated in vitro has been effective in the treatment and prevention of EBV lymphomas complicating allogeneic HSCT. However, application has been limited by the lack of specialized facilities required for T-cell production and the time required (approximately 65 days) to generate EBV-specific T-cells that are adequate in number, EBV-specificity and lack of alloreactivity. Effective treatment with EBV-specific T-cells may also be hampered or prevented by limited availability of donor cells for T-cell propagation, lack of prior sensitization of the donor to the virus in vivo, or failure of the donor's T-cells to recognize EBV in the context of an HLA allele expressed by the tumor. Based on our preliminary studies, we hypothesize that EBV-specific T-cells derived from appropriately selected third party donors from a pre established bank of T-cell lines can address these limitations. Accordingly, in Aim 1 we will: conduct a Phase II trial of EBV-specific T-cells derived from third party donors selected on the basis of matching for 2 HLA alleles and appropriate HLA restriction, in the treatment of patients with EBV lymphomas complicating allogeneic HSCT or organ transplants or other EBV-associated malignancies associated with immune deficiencies. In this trial, we will correlate clinical responses and alterations in circulating levels of EBV DNA observed with quantitative evaluations of the in vivo growth, expansion and survival of the EBV-specific T-cells in vivo following adoptive transfer. In Aim 2, we will establish of a large, genetically diverse bank of EBV-specific T-cell lines specifically consented for third party use. Each of the 300 T-cell lines that has been previously generated from consenting donors, and is a candidate line for this bank has been manufactured under GMP conditions and characterized as to its HLA genotype, EBV specificity, lack of alloreactivity and requisite microbial sterility. We plan to identify the HLA restriction of the EBV-specific T-cells in each line, so as to permit selection of EBV T-cells for a given patient that are restricted by an HLA allele shared by the patient's tumor. We will also identify lines containing T-cells specific for LMP-1 and/or LMP-2, which could be used to treat EBV+ malignancies that selectively express these antigens. We expect this phase II trial will provide the data needed to initiate a phase III study comparing this therapy with chemotherapy and/or Rituxan. The bank of well characterized EBV-specific T-cells should also provide a broadly applicable and accessible resource for such trials.