POST TRANSPLANT T CELL RECONSTITUTION IN NEUROBLASTOMA

描述(申请人提供)：我们的长期目标是开发联合免疫疗法和生物疗法，以提高高危神经母细胞瘤患者的存活率。神经母细胞瘤(NB)是儿童期最常见的颅外实体瘤，长期存活率低。为了改善这些结果，我们最近进行了一项试验，以确定联合大剂量化疗和自体干细胞挽救的可行性。我们发现令人振奋的3年EFS为58%，然而免疫重建受损是该治疗方案的主要发病率。在这项试验中，我们建议评估串联移植后患者细胞免疫重建的可行性。我们将仔细评估输注共刺激扩增的自体T细胞(T细胞增强；TCA)对免疫恢复的影响，为设计自体治疗性细胞神经母细胞瘤疫苗奠定基础。针对PAR-03-005，这项建议的具体目的是：(1)进行两部分临床试验，研究高危NB患儿移植后TCA的可行性，并测试TCA对免疫重建的影响。在第一部分，我们将完成正在进行的TCA在12名高危NBL患者中的先导性研究。在第二部分中，我们将在另外24名患者中进行TCA的随机II期试验，测试TCA对CD4重建和结合疫苗应答的影响。我们将确定(A)体外扩增的T细胞群体中肿瘤污染率；(B)过继转移是否增加T细胞恢复率和疫苗反应；(C)临床毒性；(D)在这一高危患者组中的EFS。(2)(A)通过测量(I)通过流式细胞仪检测外周血T淋巴细胞亚群，(Ii)通过分型检测CD4+T淋巴细胞受体谱，(Iii)临床DTH对召回抗原的反应，以及(Iv)对结合疫苗的反应，来确定接受SCR和T细胞增强的患者T细胞重建的速度和质量。(B)使用这些参数来比较接受T细胞增强与标准SCR的患者。(3)评估(I)接受SCR和TCA的患者的抗NB免疫，以及(Ii)评估Survivin在NB中的潜在肿瘤抗原。所使用的检测方法包括：ELISPOT中对装载肿瘤RNA、存活RNA或对照RNA的靶点的干扰素-伽马的产生，通过多肽/MHC四聚体在人类白细胞抗原A2+患者中产生的Survivin特异性细胞毒性T淋巴细胞，以及对装载肿瘤、Survivin或对照RNA的靶标的CTL反应。

DESCRIPTION (provided by applicant): Our long-term goal is to develop combined immunotherapy and biologic therapies to improve the survival of patients with high-risk neuroblastoma. Neuroblastoma (NB) is the most common extracranial solid tumor of childhood, with a poor long-term survival rate. In an effort to improve these results, we recently performed trial to determine the feasibility of tandem high-dose chemotherapy with autologous stem cell rescue. We found an encouraging 3-year EFS of 58%, however impaired immune reconstitution constituted the major morbidity of this treatment regimen. In this trial we propose to assess the feasibility of cellular immune reconstitution in patients following tandem transplantation. We will carefully assess the effect of infusion of co-stimulated expanded autologous T cells (T cell augmentation; TCA) on immune recovery, establishing the infrastructure to engineer autologous therapeutic cellular neuroblastoma vaccines. In response to PAR-03-005, the SPECIFIC AIMS of this proposal are to: (1) Conduct a two-part clinical trial to study the feasibility of post-transplant TCA in pediatric patients with high-risk NB and test the impact of TCA on immune reconstitution. In the first part, we will complete the ongoing pilot study of TCA in high-risk NBL in 12 patients. In the second part we will perform a randomized phase II trial of TCA in additional 24 patients, testing the impact of TCA on CD4 reconstitution and conjugate vaccine response. We will determine (a) the rate of tumor contamination in ex vivo expanded T cell populations; (b) whether adoptive transfers increase the rate of T cell recovery and vaccine response; (c) clinical toxicity; and (d) the EFS in this high-risk group of patients. (2) (A) Determine the pace and quality of T cell reconstitution for patients undergoing SCR and T cell augmentation by measuring: (i) peripheral blood T lymphocyte subsets via flow cytometry, (ii) CD4+ T lymphocyte receptor repertoire via spectratyping, (iii) clinical DTH reactions to recall antigen, and (iv) response to a conjugate vaccine. (B) Use these parameters to compare patients receiving T cell augmentation vs. standard SCR. (3) Assess (i) anti-NB immunity for patients undergoing SCR and TCA and (ii) assess survivin as a potential tumor antigen in NB. Assays used will include: IFN- gamma production in ELISPOT assays to targets loaded with tumor RNA, surviving RNA, or control RNA, survivin-specific cytotoxic T lymphocytes via peptide/MHC tetramers in HLA-A2+ patients, and CTL response to targets loaded with tumor, survivin, or control RNA.