The role of TRAIL in bone marrow transplantation

描述(申请人提供)：捐献者T细胞在 异基因骨髓移植。供者来源的细胞毒T细胞 主要负责移植物抗肿瘤(GVT)活动和 移植物抗宿主病(GVHD)细胞毒T细胞可以发挥其细胞溶解作用 至少通过两条效应通路发挥活性：Fas配体(FasL)和 穿孔素/颗粒酶。这两个细胞毒效应通路，除了肿瘤坏死因子 (已被认为是第三种T细胞效应途径)，发挥着重要的作用 在移植物抗宿主病和移植物抗宿主病的发展中的作用。我们在小鼠模型方面的最新研究 而其他人则通过以下方式展示了这些细胞溶解途径的不同用途 供者T细胞用于GVHD、GVT和供者白细胞输注(DLI)。这表明 选择性地抑制细胞溶解途径可能代表着一种新的 将GVT与GVHD分离的策略。肿瘤坏死因子相关 细胞凋亡诱导配体(TRAIL)是肿瘤坏死因子超家族的成员之一。 包括FasL和肿瘤坏死因子。TRAIL已被证明可以诱导肿瘤细胞的凋亡 在不影响未转化细胞的情况下，然而它在健康和疾病中的作用 在很大程度上仍不为人所知。在本申请中，我们建议剖析 使用最近生成的GVHD、GVT和供体植入y的试验 TRAIL缺陷小鼠与临床中和抗TRAIL抗体 相关的小鼠骨髓移植模型。在具体目标1中，我们建议研究这些影响 A)TRAIL缺陷的B6(B6-/-)供体细胞，或b)体内 给予中和抗TRAIL抗体对移植物抗宿主病发病率和 死亡率和特定的移植物抗宿主病相关器官病理。我们将研究这一角色 供者分析TRAIL在移植物抗宿主病病理生理学中的作用 T细胞扩增和活性、CD4/8比值、血清细胞因子水平(IL-1、IL-12、 肿瘤坏死因子、干扰素)、血清内毒素水平和巨噬细胞活化。这一分析 将对a)已收到亚致死剂量的收件人重复 确定移植物排斥反应的辐射，或b)接种了肿瘤 分析TRAIL通路在骨髓移植时对GVT的重要性 (和DLI)与GVHD有关。在具体目标2中，我们将研究 受体TRAIL缺陷及其对移植物抗宿主病发生的影响 在具体目标3中，我们建议产生具有多个缺陷的小鼠 细胞溶解途径。这些小鼠将用于类似的实验 在目标1和2中描述，以发现任何冗余、协同或对抗 这些在移植物抗宿主病、移植物抗宿主病和植入中的细胞溶解途径。最后，具体地说 目的4我们提出通过TRAIL的过表达来增强GVT的活性 (通过腺病毒介导的传递)在供者T细胞中。这些研究可以提供 更好地理解TRAIL通路在(A)T中的作用 体内细胞毒性，以及(B)移植物抗宿主病、移植物抗宿主病和骨髓移植后的植入，其中 可以立即在临床上应用。

DESCRIPTION (provided by applicant): The donor T cell plays a pivotal role in allogeneic bone marrow transplantation (BMT). Cytotoxic T cells of donor origin are primarily responsible for graft-versus-tumor (GVT) activity and graft-versus-host-disease (GVHD). Cytotoxic T cells can exert their cytolytic activity through at least two effector pathways: Fas ligand (FasL) and perforin/granzyme. These two cytotoxic effector pathways, in addition to TNF (which has been proposed as a third T cell effector pathway), play an important role in the development of GVHD and GVT. Recent studies in murine models by us and others have demonstrated a differential use of these cytolytic pathways by donor T cells in GVHD, GVT and donor leukocyte infusion (DLI). This suggests that the selective inhibition of a cytolytic pathway could represent a novel strategy for the separation of GVT from GVHD. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of the TNF superfamily, which includes FasL and TNF. TRAIL has been shown to induce apoptosis of tumor cells without affecting nontransformed cells, however its role in health and disease is still largely unknown. We propose in this application to dissect the role of TRAIL in GVHD, GVT, and donor engraftment y using recently generated TRAIL-deficient mice and neutralizing anti-TRAIL antibodies in clinically relevant murine BMT models. In Specific Aim 1 we propose to study the effects of a) TRAIL-deficient B6 (B6.TRAIL-/-) donor cells, or b) in vivo administration of a neutralizing anti-TRAIL antibody on GVHD morbidity and mortality and specific GVHD-associated organ pathology. We will study the role of TRAIL in the pathophysiology of GVHD in the recipients by analysis of donor T cell expansion and activity, CD4/8 ratio, serum cytokine levels (IL-1, IL-12, TNF, IFN), serum endotoxin levels, and macrophage activation. This analysis will be repeated with recipients which either a) have received sublethal irradiation to determine graft rejection, or b) have been inoculated with tumor cells at the time of BMT to analyze the importance of the TRAIL pathway for GVT (and DLI) in relation to GVHD. In Specific Aim 2 we will study the effects of TRAIL-deficiency in the recipient and its effects on the development of GVHD. In Specific Aim 3 we propose to generate mice with multiple deficiencies in the cytolytic pathways. These mice will be used for similar experiments as described in Aims 1 and 2 to discover any redundancy, synergy or antagonism of these cytolytic pathways in GVHD, GVT, and engraftment. Finally, in Specific Aim 4 we propose to enhance GVT activity through the overexpression of TRAIL (by adenovirus-mediated delivery) in donor T cells. These studies could provide a better mechanistic understanding of the role of the TRAIL pathway in (a) T cell cytotoxicity in vivo, and (b) GVHD, GVT and engraftment after BMT, which could have immediate clinical applicability.