NK Cells and Their Receptor in Leukemia Therapy

在当前的资助期间，我们通过在晚期AML患者中实现完全缓解，证明了过继转移的成人NK细胞的显著抗白血病潜力。该项目将包括新的临床试验，旨在改善利用脐带血（UCB）移植中NK细胞同种异体反应性的技术，以及体外和小鼠实验，以表征支持成人血液和UCB祖细胞NK细胞扩增和培养的机制。在SA 1中，我们将进行一系列临床试验，评估IL-2给药在成人NK细胞过继转移/T细胞耗竭的UCB移植策略中增强体内NK细胞扩增和教育的疗效。我们已经产生了体外数据，表明IL-15对NK细胞扩增和分化的重要作用。临床级IL-15最近已由NCI提供给我们，我们接下来将在第一次人体试验中用IL-15代替IL-2，以测试IL-15促进UCB移植后移植NK细胞的教育的能力。第三项临床试验将检验过继转移>10倍的假设。 较高剂量的离体扩增的成体NK细胞将进一步增强功效。在SA 2中，我们将监测暴露于IL-2或IL-15的患者中、项目1（瓦格纳）中UCB移植试验的Treg输注后以及对病毒应激有反应的移植受者中扩增的NK细胞的受体库、功能和教育。我们还将使用临床前体外试验和小鼠模型，其中成年人NK细胞和UCB祖细胞被转移到NOD SCID IL 2 Rgamma-c-null免疫缺陷小鼠中，以测试增强NK细胞功能的新策略。具体而言，我们将测试增强NK细胞抗白血病功能的方法，并有可能转化为临床试验。我们将测试使用IL-15 Ra-Fc的IL-15的反式呈递，使用IL-15 Ra+人工K562细胞刺激细胞（Notch配体或41 BB配体）的离体NK细胞扩增，以及通过与表达单个Bw 4、C1或C2 KIR配体的K562刺激物一起培养来定制离体扩增的NK细胞的同种异体反应特异性的技术。最终，来自这一系列旨在优化NK细胞抗白血病作用的临床试验的最佳策略将与项目1和项目2中开发的增强免疫重建和最小化GVHD的临床方法相结合。 项目2.

During the current funding period we demonstrated the significant anti-leukemic potential of adoptively transferred adult NK cells by effecting complete remissions in patients with advanced AML. This project will include novel clinical trials designed to improve techniques exploiting NK cell alloreactivity in umbilical cord blood (UCB) transplants as well as in vitro and murine experiments to characterize the mechanisms supporting the expansion and education of NK cells derived from adult blood and from UCB progenitors. In SA1, we will perform a series of clinical trials assessing the efficacy of IL-2 administration to enhance in vivo NK cell expansion and education in an adult NK cell adoptive transfer/T cell depleted UCB transplantation strategy. We have generated in vitro data suggesting an important effect of IL-15 on NK cell expansion and differentiation. Clinical grade IL-15 has recently been made available to us by the NCI, and we will next substitute IL-15 for IL-2 in the first human trial to test the ability of IL-15 to facilitate education of engrafting NK cells after UCB transplant. A third clinical trial will test the hypothesis that adoptive transfer of >10 fold higher doses of ex vivo expanded adult NK cells will further enhance efficacy. In SA2 we will monitor the receptor repertoires, function and education of NK cells expanding in patients exposed to IL-2 or IL-15, after Treg infusion from the UCB transplant trials in Project 1 (Wagner), and in transplant recipients responding to viral stress. We will also use pre-clincial in vitro assays and a murine model in which adult human NK cells and UCB progenitors are transferred into NOD SCID IL2Rgamma-c-null immunodeficient mice to test novel strategies to enhance NK cell function. Specifically, we will test methods to enhance the anti-leukemic function of NK cells with potential for translation into clinical trials. We will test trans-presentation of IL-15 using lL-15Ra-Fc, ex vivo NK cell expansion using IL-15Ra+ artificial K562 cell stimulator cells (¿ Notch ligand or41BB ligand), and techniques to tailor the allo-reactive specificity of ex vivo expanded NK cells by culture with K562 stimulators expressing single Bw4, C1 or C2 KIR ligands. Ultimately, the best strategy from this series of clinical trials designed to optimize the anti-leukemic effect of NK cells will be combined with the clinical methods to enhance immune reconstitution and minimize GVHD developed in Project 1 and in Project 2.