GENETICS AND MECHANISMS OF DLI EFFECTS FOLLOWING NON-MYELOBLATIVE HCT

非清髓性造血细胞移植(HCT)后延迟输注供者白细胞 (DLI)是一种很有前途的免疫治疗方法，用于治疗包括白血病和 淋巴瘤。然而，这种方法的主要局限性包括移植物失败、移植物抗宿主的风险。 疾病、移植物抗宿主病和感染。MGH-MHC近交系小型猪提供了一种临床前模型 移植生物学，对红细胞压积的反应与人类相似。初步数据显示， 新的最小限度骨髓抑制制备方案在大剂量后导致稳定的多系嵌合体 单倍体相合的Hct，不会引起GVHD。在这项建议中，我们的目标是1)确定免疫学上的 参与控制hbst抗移植物(HVG)和GVH反应的机制，从而允许植入 在这个模型中，没有GVHD跨越MHC障碍。这些研究将与以下机构密切合作 项目1和项目2扩展了啮齿动物模型的机理研究。然后我们会对猪进行优化 模型，以便于将该方案翻译到临床并分析特定基因的重要性 差异(单倍体完全相同的第I类和第II类，仅适用于第I类，仅适用于第II类)植骨、GVHD和 随后的DLI。在与项目4的合作下，我们将测试改进干细胞收获的新策略 通过甲状旁腺激素(PTH)刺激进行细胞因子动员。增加了茎的数量 甲状旁腺素刺激和细胞因子动员后的白细胞分离产物中的细胞可以使稳定 使用较低剂量的细胞进行植入在临床上更容易实现。希望这些工作的结果 研究将允许开发在嵌合患者中使用DLI的量身定制的方法，这取决于 关于他们与捐献者之间的人类白细胞抗原差异。此外，我们计划进一步开发猪模型，以允许 直接评估HCT和DLI的移植物抗肿瘤效应。为此，我们将建立肿瘤细胞 从近亲交配的小型猪衍生的肿瘤株，并使这些肿瘤株适应猪的体内生长。与 最近可获得组织相容的小型猪和由这些高度近交系衍生的肿瘤系 对于动物，我们有独特的机会在临床前的大型动物模型中开发可移植的肿瘤。 这些研究可能为未来免疫治疗方法的治疗提供基础。 血液学恶性疾病，可转化为治疗人类疾病的疾病。

Non-myeloablative hematopoietic cell transplantation (HCT) followed by delayed donor leukocyte infusion (DLI) is a promising immunotherapeutic approach to treat hematologic malignancies including leukemias and lymphomas. Major limitations of this approach, however, include the risks of graft failure, graft-versus-host disease, (GVHD) and infection. MGH MHC-inbred miniature swine provide a pre-clinical model for studies of transplantation biology with responses to HCT resembling those of humans. Preliminary data suggest that a novel, minimally myelosuppressive preparative regimen leads to stable multilineage chimerism following highdose haploidentical HCT, without causing GVHD. In this proposal we aim to 1) determine the immunological mechanisms involved in controlling hbst-versus-graft (HVG) and GVH responses that allow engraftment without GVHD across MHC barriers in this model. These studies will be performed in close collaboration with Projects 1 and 2 to extend the mechanistic studies in rodent models. We will then 2) optimize the swine model to facilitate translation of this protocol to the clinic and analyze the importance of specific genetic disparities (haploidentical class I and II, class I only, class II only) on engraftment, GVHD and the effects of subsequent DLI. In collaboration with Project 4, we will test novel strategies to improve stem cell harvests following cytokine mobilization through parathyroid hormone (PTH) stimulation. Increased numbers of stem cells in the leukapheresis product following PTH stimulation and cytokine mobilization may enable stable engraftment using lower doses of cells more easily attainable in the clinic. It is hoped that results of these studies will permit the development of tailored approaches to the use of DLI in chimeric patients depending on their HLA disparities from the donor. In addition, we plan to 3) further develop the swine model to allow direct assessment of graft-versus-tumor effects of HCT and DLI. For this purpose, we will establish tumor cell lines derived from inbred miniature swine and adapt these tumor lines for in vivo growth in pigs. With the recent availability of histocompatible miniature swine, and tumor lines derived from these highly inbred animals, we have the unique opportunity to develop transplantable tumors in a preclinical large animal model. These studies could provide a foundation for future immunotherapeutic approaches for the treatment of hematological malignancies that may be translated toward treatment of human disease.