Enhancing GvL via delayed ex-vivo co-stimulated DLI after non-myeloablative SCT

非清髓性 SCT 后通过延迟离体共刺激 DLI 增强 GvL

• 批准号: 7295714
• 负责人: STEVEN C GOLDSTEIN
• 金额: $ 27.15万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-29 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7295714
• 关键词: 5-(6)-carboxyfluorescein diacetate succinimidyl ester; Acute leukemia; Adoptive Immunotherapy; Adult; Allogenic; Antigens; Attenuated; Autoantigens; Biological Assay; Cell physiology; Cells; Chimerism; Clinical; Cytomegalovirus; Disease; Dysmyelopoietic Syndromes; End Point; Foundations; Goals; Hematologic Neoplasms; Homologous Transplantation; Human Herpesvirus 4; Immune; Immunophenotyping; Incidence; Inflammatory; Infusion procedures; Interferon Type II; Life; Lymphocyte Subset; Malignant Neoplasms; Modality; Numbers; Opportunistic Infections; Organ; Outcome; Patients; Pennsylvania; Phase I Clinical Trials; Reading; Relapse; Residual Neoplasm; Risk; Safety; Severities; Spiral Computed Tomography; Staphylococcal Enterotoxin B; Stem cell transplant; Stimulus; T-Lymphocyte; Tetradecanoylphorbol Acetate; Time; Toxic effect; Translational Research; Transplantation; Universities; Work; base; cytokine; cytotoxicity; day; enzyme linked immunospot assay; experience; graft vs host disease; graft vs leukemia effect; improved; killings; leukemia; novel; novel strategies; pilot trial; prophylactic; reconstitution; response; tumor

DESCRIPTION (provided by applicant): The long-term goal of this project is to establish a novel clinical platform for enhancing immune reconstitution, and more specifically, the graft vs leukemia (GvL) effect post-transplant via prophylactic infusion of ex-vivo co-stimulated allogeneic DLI to improve the outcome for patients with acute leukemia and myelodysplastic syndrome. This strategy is based on our hypothesis that activated donor T-cells given after donor chimerism is established, at a time of minimal residual disease (MRD), and in the absence of the inflammatory milieu of the early post-transplant period, will induce a more potent GvL effect without causing severe GvHD. This study will provide a foundation for efforts to implement tumor-specific adoptive cellular therapy, i.e., enhancing GvL without increasing GvHD. Specific Aim 1: Conduct a phase I clinical trial to confirm the feasibility and safety of delayed infusion of activated DLI after T-cell depleted non-myeloablative allogeneic stem cell transplantation in adult patients with high risk hematologic malignancies. The incidence and severity of graft vs host disease will be a primary endpoint. Specific Aim 2: Assess the impact of prophylactic activated DLI on tumor-specific cytotoxicity and immune reconstitution: 2a) Study T-cell responses to leukemia-specific antigen (autologous tumor), polyclonal stimuli (SEB, PMA), and recall antigens (CMV, EBV), to assess for the enhanced immune potential of donor T-cells before and after ex-vivo costimulation via functional analysis of donor T-cell function based on 3 complementary read-outs: (i) proliferation via CFSE flow cytometric assay, (ii) cytokine secretion via interferon gamma release (ELISPOT), and (iii) cytotoxicity via degranulation assay as a marker of specific target killing based on flow cytometric analysis of CD107a. 2b) Study T-cell proliferative responses (proliferation, cytokine secretion, cytotoxicity) to leukemia-specific antigen (autologous tumor), polyclonal stimuli (SEB, PMA), and recall antigens (CMV, EBV), to assess for enhanced immune reconstitution of recipient T-cells before and after infusions of ex-vivo costimulated donor cells via functional analysis of recipient T-cells using the assays outlined above at 5 time points; pre- transplant, approximately day+90 (pre pADLI #1), approximately day+160 (pre pADLI #2), day+270, and day+365.

描述（申请人提供）：该项目的长期目标是建立一个新的临床平台，用于增强免疫重建，更具体地说，通过预防性输注离体共刺激同种异体 DLI 来增强移植后的移植物抗白血病（GvL）效应，以改善急性白血病和骨髓增生异常综合征患者的预后。该策略基于我们的假设，即在供体嵌合建立后、在微小残留病 (MRD) 时以及在移植后早期不存在炎症环境的情况下，给予活化的供体 T 细胞，将诱导更有效的 GvL 效应，而不会引起严重的 GvHD。这项研究将为实施肿瘤特异性过继细胞疗法（即在不增加 GvHD 的情况下增强 GvL）的努力奠定基础。具体目标 1：开展 I 期临床试验，以确认高危血液恶性肿瘤成年患者在 T 细胞耗尽的非清髓性同种异体干细胞移植后延迟输注活化 DLI 的可行性和安全性。移植物抗宿主病的发生率和严重程度将是主要终点。具体目标 2：评估预防性激活 DLI 对肿瘤特异性细胞毒性和免疫重建的影响：2a) 研究 T 细胞对白血病特异性抗原（自体肿瘤）、多克隆刺激（SEB、PMA）和回忆抗原（CMV、EBV）的反应，以评估供体 T 细胞在离体前后增强的免疫潜力 基于 3 个互补读数，通过对供体 T 细胞功能进行功能分析进行共刺激：(i) 通过 CFSE 流式细胞术测定进行增殖，(ii) 通过干扰素 γ 释放 (ELISPOT) 进行细胞因子分泌，以及 (iii) 通过脱颗粒测定进行细胞毒性，作为基于 CD107a 的流式细胞术分析的特异性靶标杀伤标记。 2b) 研究 T 细胞对白血病特异性抗原（自体肿瘤）、多克隆刺激（SEB、PMA）和回忆抗原（CMV、EBV）的增殖反应（增殖、细胞因子分泌、细胞毒性），以评估通过功能性体外共刺激供体细胞输注前后受体 T 细胞的免疫重建增强情况 使用上述测定法在 5 个时间点对受体 T 细胞进行分析；移植前，大约+90天（pADLI#1前）、大约+160天（pADLI#2前）、+270天和+365天。

项目成果

Efficient clinical-scale enrichment of lymphocytes for use in adoptive immunotherapy using a modified counterflow centrifugal elutriation program.

使用改良的逆流离心淘洗程序，有效地进行临床规模的淋巴细胞富集，用于过继性免疫治疗。

• DOI: 10.3109/14653240903188921
• 发表时间: 2009
• 期刊: Cytotherapy
• 影响因子: 4.5
• 作者: PowellJr,DanielJ;Brennan,AndreaL;Zheng,Zhaohui;Huynh,Hong;Cotte,Julio;Levine,BruceL
• 通讯作者: Levine,BruceL