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

描述(申请人提供)：EB病毒引起的淋巴增殖性疾病和EBV+淋巴瘤是异基因造血细胞(HSCT)和器官移植(SOT)受者以及患有严重遗传和获得性免疫缺陷患者的发病率和死亡率的重要原因。EBV+霍奇金氏病(HD)、EBV+噬血细胞淋巴组织细胞增生症、EBV+鼻咽癌和EBV+平滑肌肉瘤也构成了一组恶性肿瘤，目前的化疗往往无效。体外产生的供者来源的EBV特异性T细胞过继治疗在治疗和预防EBV淋巴瘤合并异基因HSCT方面是有效的。然而，由于缺乏生产T细胞所需的专门设施，以及产生数量充足、EBV特异性和缺乏同种异体反应的EBV特异性T细胞所需的时间(约65天)，应用一直受到限制。对EBV特异性T细胞的有效治疗也可能受到以下因素的阻碍或阻止：供体细胞用于T细胞繁殖的可获得性有限，供体缺乏体内对病毒的事先敏感性，或供体的T细胞未能识别肿瘤表达的HLA等位基因中的EBV。基于我们的初步研究，我们假设，从预先建立的T细胞库中适当选择第三方捐赠者的EBV特异性T细胞可以解决这些限制。因此，在目标1中，我们将进行第二阶段试验，对来自第三方捐赠者的EBV特异性T细胞进行第二阶段试验，这些T细胞来自于根据2个HLA等位基因匹配和适当的HLA限制而选择的，用于治疗EBV淋巴瘤合并异基因HSCT或器官移植或其他与免疫缺陷相关的EBV相关恶性肿瘤。在这项试验中，我们将观察到的临床反应和循环中EBV DNA水平的变化与过继转移后EBV特异性T细胞在体内的生长、扩增和存活的定量评估相关联。在目标2中，我们将建立一个大型的、基因多样化的EBV特异性T细胞系库，专门同意第三方使用。先前从自愿捐赠者那里获得的300个T细胞系中的每一个都是在GMP条件下生产的，其特征是其HLA基因、EBV特异性、缺乏同种异体反应性和必要的微生物无菌。我们计划确定每个品系中EBV特异性T细胞的HLA限制性，以便为给定的患者选择受患者肿瘤共享的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细胞的株，这些T细胞可用于治疗选择性表达这些抗原的EBV+恶性肿瘤。我们预计这项II期试验将提供启动III期研究所需的数据，将这种疗法与化疗和/或Rituxan进行比较。具有良好特性的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。 公共卫生相关性：EBV相关的PTLDS和淋巴瘤是接受造血细胞和实体器官移植的患者以及患有严重遗传和获得性免疫缺陷的患者的主要和危及生命的并发症。目前采用化疗和/或利妥昔单抗的治疗方法在不超过50%的病例中持续有效，并可能导致严重的毒性反应。过继转移供者来源的EBV特异性T细胞被发现对预防和治疗这些疾病是有效的。在英国和我们中心的学习也可能是有效的。这项拨款建议对第三方来源的EBV特异性T细胞进行第二阶段试验，以评估这一方法。该T细胞是根据部分匹配和肿瘤导向的人类白细胞抗原限制而选择的。它还建议建立一个完全特化的、经特别同意的GMP级EBV特异性T细胞库，以便为这些疾病的采用治疗提供一个广泛适用和立即可用的资源。

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. PUBLIC HEALTH RELEVANCE: EBV-associated PTLDs and lymphomas are a major and life-threatening complication for patients receiving hematopoietic cell and solid organ transplants and patients with severe genetic and acquired immunodeficiencies. Current treatment with chemotherapy and/or Rituxan is durably effective in no more than 50% of cases and may induce significant toxicity. Adoptive transfer of donor-derived EBV specific T-cells has been found to be effective for prevention and treatment of these disorders. Studies in the United Kingdom and at our center may also be effective. This grant proposes a Phase II trial of third-party derived EBV-specific T-cells selected on the basis of partial HLA matching and tumor-directed HLA restriction to assess this approach. It also proposes the establishment of a fully characterized specifically consented bank of GMP-grade EBV-specific T-cells so as to provide a broadly applicable and immediately accessible resource for adoptive therapy of these diseases.