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

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，以增强免疫重建，更具体地说，移植物抗白血病(GVL)通过预防性输注体外共刺激同种异体DLI来影响移植后的效果，以改善急性白血病和骨髓增生异常综合征患者的预后。这一策略是基于我们的假设，即在供者嵌合体建立后，在微小残留病(MRD)时期，在没有移植后早期炎症环境的情况下，在不引起严重GvHD的情况下，给予激活的供者T细胞将产生更有效的GVL效应。这项研究将为实施肿瘤特异性过继细胞治疗，即在不增加GvHD的情况下增强GVL奠定基础。具体目的1：进行I期临床试验，以确定成人高危恶性血液病患者T细胞耗竭的非清髓性异基因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。具体目的2：评估预防性激活的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)和召回抗原(CMV、EBV)的反应，通过基于3种互补读数的供体T细胞功能分析来评估体外共刺激前后供体T细胞的免疫潜力增强：(I)CFSE流式细胞仪检测增殖；(Ii)通过干扰素-γ释放(ELISPOT)分泌细胞因子；以及(Iii)基于CD107a的流式细胞术分析，通过脱颗粒试验作为特异性靶向杀伤的标志的细胞毒性。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)和召回抗原(CMV、EBV)的增殖反应(增殖、细胞因子分泌、细胞毒性)，以评估输注体外共刺激供体细胞前后受者T细胞的增强免疫重建，通过使用上述5个时间点的检测对受者T细胞进行功能分析；移植前，大约+90天(Padli#1前)，大约+160天(Padli#2前)，+270天和+365天。

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.