Genetic immunotherapy for malignancy: murine modeling

描述（由申请人提供）：免疫疗法作为恶性肿瘤的第四种治疗方式具有很大的前景，但仍然存在重大障碍，包括肿瘤特异性，维持效应T细胞功能和最佳激活策略。我们已经开发了一种用抗CD 3和抗CD 28激活原代T细胞，然后进行逆转录病毒转导的系统，以产生表达基因工程肿瘤特异性受体的T细胞。虽然初步实验显示出治疗前景，但这些细胞在体内的动力学、功能和IL-2依赖性尚未得到严格研究。基因治疗修饰的表达新的肿瘤特异性α/β T细胞受体（TCR）的原代T细胞在体内的功效仍有待证实。卵清蛋白已被用作许多系统中的替代肿瘤抗原。我们建议创建一个双顺反子逆转录病毒表达载体编码卵清蛋白特异性T细胞受体α和β基因的OT-1 T细胞受体转基因小鼠（OT-1 Tg）。我们将使用Ly 5.1同源供体比较基因治疗修饰的T细胞与OT-1 Tg T细胞在过继转移后的动力学、IL-2依赖性和抗原应答。使用小鼠淋巴瘤模型细胞系EL-4和小鼠白血病模型细胞系C1498，每个细胞系都被转导以表达卵清蛋白，我们将测试基因治疗修饰的T细胞的治疗功效。这些研究将提供重要的数据，用于模拟未来的遗传修饰T细胞过继免疫疗法治疗淋巴瘤和白血病的临床试验。

DESCRIPTION (provided by applicant): Adoptive immunotherapy has great promise as a fourth treatment modality for malignancy, but significant obstacles remain, including tumor specificity, maintenance of effector T cell function and optimal activation strategy. We have developed a system of primary T cell activation with anti-CD3 and anti-CD28 followed by retroviral transduction to generate T cells expressing genetically engineered, tumor-specific receptors. While preliminary experiments show therapeutic promise, the kinetics, function and IL-2 dependence of such cells in vivo has not been rigorously studied. The efficacy of gene-therapy modified primary T cells expressing a new, tumor-specific alpha/beta T cell receptor (TCR) in vivo remains to be demonstrated. Ovalbumin has been used as a surrogate tumor antigen in many systems. We propose to create a bicistronic retroviral expression vector encoding the ovalbumin-specific T cell receptor alpha and beta genes from the OT-1 T cell receptor transgenic mouse (OT-1 Tg). We will use Ly 5.1 congenic donors to compare gene-therapy modified T cells to OT-1 Tg T cells for kinetics, IL-2 dependence and antigen response after adoptive transfer. Using the murine lymphoma model cell line EL-4 and the murine leukemia model cell line C 1498, each transduced to express ovalbumin, we will test the therapeutic efficacy of gene-therapy modified T cells. These studies will provide data important for modeling future clinical trials of adoptive immunotherapy with genetically modified T cells to treat lymphoma and leukemia.