ENHANCED IMMUNOGENICITY OF NEUROBLASTOMA CELLS BY GENETIC ENGINEERING

神经母细胞瘤是儿童最常见的颅外实体瘤。 尽管有能力实现最小残留病(MRD) 清髓治疗和自体骨髓移植(ABMT)， 超过50%的高危患者会出现反复发作的致命疾病。我们希望 确定神经母细胞瘤细胞是否进行主动特异性免疫治疗 被设计成唤起免疫反应的基因可以在根除 MRD和改善高危患者的预后。我们将介绍 细胞因子基因，特别是干扰素-γ和粒-巨噬细胞集落刺激因子基因； 转化为自体和人类白细胞抗原单基因匹配的同种异体神经母细胞瘤 细胞，检测细胞因子的表达水平，并确定表型 改变。一组细胞因子转导的肿瘤细胞稳定表达 将衍生出共同的HLAA或B抗原。神经母细胞瘤的能力 处理和呈递HTLV-1特异性多肽抗原的细胞 将测定TAX和MAGE-1作为细胞溶解T细胞(CTL)的靶点。这个 细胞因子表达自体和MHC-I类分子的能力匹配 同种异体神经母细胞瘤细胞诱导CTL的评估如下 给患者注射基因工程肿瘤细胞。 使用逆转录病毒干扰素-γ生产载体的初步结果是 足够鼓舞人心，足以保证进行1期/磅临床试验。未来 使用GM-CSF转导的神经母细胞瘤细胞的临床试验 想象中的。神经母细胞瘤提供了一种独特的检测环境 基于细胞可获得性的免疫治疗基因转移策略 LINES，尽管达到了MRD状态，可预测的复发率，以及 细胞系的生长与低存活率有很强的相关性。

Neuroblastoma is the most common extracranial solid tumor in children. Despite the ability to achieve minimal residual disease (MRD) with myeloablative therapy and autologous bone marrow transplantation (ABMT), over 50% of high risk patients develop recurrent fatal disease. We wish to determine whether active specific immunotherapy with neuroblastoma cells engineered to evoke an immune response can play a role in eradication of MRD and improve prognosis for high risk patients. We shall introduce cytokine genes, specifically interferon-gamma (IFN-gamma) and GM-CSF genes, into autologous and HLA single locus matched allogeneic neuroblastoma cells, assay levels of cytokine expression, and determine phenotypic changes. A panel of cytokine transduced tumor cells that stably express common HLA A or B antigens will be derived. The ability of neuroblastoma cells to process and present specific peptide antigens derived from HTLV-1 tax and MAGE-1 as targets for cytolytic T-cells (CTL) will be assayed. The ability of cytokine expressing autologous and MHC class I matched allogeneic neuroblastoma cells to elicit CTL will be assessed following administration of genetically engineered tumor cells to patients. Preliminary results using retroviral IFN-gamma producing vectors are sufficiently encouraging to warrant a phase 1/lb clinical trial. Future clinical trials employing GM-CSF transduced neuroblastoma cells are envisioned. Neuroblastoma offers a unique setting in which to test immunotherapeutic gene transfer strategies due to availability of cell lines, the predictable relapse rate despite achievement of MRD status, and the strong correlation of cell line outgrowth with poor survival.