IMMUNE RESPONSES INDUCED BY GENE TRANSFER

基因转移引起的免疫反应

• 批准号: 6102908
• 负责人: ALFRED E CHANG
• 金额: $ 18.1万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-06-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6102908
• 关键词: CD3 molecule; antibody specificity; cellular immunity; clinical research; cytotoxic T lymphocyte; dendritic cells; genetic transduction; histocompatibility antigens; histocompatibility gene; host neoplasm interaction; human therapy evaluation; immune response genes; immunofluorescence technique; immunosuppression; interleukin 2; laboratory mouse; neoplasm /cancer genetics; neoplasm /cancer immunology; neoplasm /cancer immunotherapy; neoplasm /cancer vaccine; neoplastic growth; outcomes research; passive transport; transfection; tumor necrosis factor alpha; vector vaccine

Based upon our studies from the previous grant period, we have been able to develop two genetic approaches to upregulate immune responsiveness to tumor. The first approach involved the in vivo gene transfer of an allogeneic MHC gene into tumors which in animal and human studies, was found to reliably result in transgene expression. More importantly, we were able to elicit enhanced antitumor reactivity within the tumor and draining lymph nodes (LN); and observed tumor regression in select patients. The second approach involved the use of autologous tumor vaccines genetically altered to secrete GM-CSF as a method to augment regional immune responses in draining LN. We are currently conducting a phase I study to evaluate the use of GM- CSF transduced tumor as a vaccine to generate immune LN cells for adoptive immunotherapy. The primary focus of this project is to develop tumor reactive T cells that can be expanded ex vivo for clinical therapy. We propose to evaluate the in vitro and in vivo tumor reactivity of TIL derived from melanoma and colorectal tumors after intralesional HLA-B7 gene transfer. In addition we plan to evaluate the specificity of antigen recognition and MHC restriction of the cellular responses induced by the gene transfer. Another aspect of this project will be the application of dendritic cells (DC) which will be employed as components of tumor vaccines to sensitize LN cells for subsequent adoptive immunotherapy. DC have the unique capacity to prime naive T cells for immune responses in vitro and in vivo. Studies of DC isolated from skin tumors have demonstrated that their functional capacity can be inhibited by immunosuppressive cytokines. This has been reversed by the presence of GM-CSF with upregulation of both B7-1 and B7-2 co-stimulatory molecules. We plan to evaluate the antitumor reactivity of LN cells primed in vivo with tumor-pulsed DC admixed with DM-CSF fibroblasts in patients with melanoma and colorectal cancer. The specific aims of this project are: 1) To evaluate the immune reactivity of TIL derived from tumors modified by direct gene transfer in vivo utilizing an allogeneic class I MHC gene, 2) to evaluate the immune reactivity of T cells primed in vivo with tumor- pulsed DC with or without the admixture of GM-CSF secreting fibroblasts, and 3) To compare the immune reactivity of TIL versus primed LN in patients being treated in Aims 1 and 2.

根据我们上一个赠款期的研究，我们一直在 能够开发两种上调免疫的遗传方法 对肿瘤的反应。 第一种方法涉及体内基因 将同种异体MHC基因转移到动物和 人类研究发现可靠地导致转基因表达。 更重要的是，我们能够引起增强的抗肿瘤反应性 在肿瘤和排水淋巴结（LN）内；并观察到肿瘤 选定患者的回归。 第二种方法涉及使用 自体肿瘤疫苗在遗传上改变为GM-CSF作为一种 增加排水中区域免疫反应的方法。 我们 目前正在进行I期研究以评估GM-的使用 CSF将肿瘤作为疫苗转导，以产生免疫LN细胞 收养免疫疗法。 该项目的主要重点是发展肿瘤反应性T细胞 可以在临床疗法的情况下扩展。 我们建议 评估TIL的体外和体内肿瘤反应性 病性HLA-B7基因后的黑色素瘤和结直肠肿瘤 转移。 此外，我们计划评估抗原的特异性 识别和MHC对由细胞反应诱导的细胞反应的限制 基因转移。 该项目的另一个方面将是 树突状细胞（DC）的应用，该细胞将被用作 肿瘤疫苗的成分使LN细胞敏感 收养免疫疗法。 DC具有幼稚的独特能力 T细胞用于体外和体内的免疫反应。 DC的研究 从皮肤肿瘤中分离出来证明其功能性 免疫抑制细胞因子可以抑制能力。 这就是 GM-CSF的存在与两者的上调相反 B7-1和B7-2共刺激分子。 我们计划评估 LN细胞在体内用肿瘤脉冲DC启动的抗肿瘤反应性 黑色素瘤患者和 结直肠癌。 该项目的具体目的是：1）评估 TIL的免疫反应性来自于直接修饰的肿瘤 利用同种异体I MHC基因的体内基因转移，2） 评估肿瘤体内启动的T细胞的免疫反应性 有或没有GM-CSF分泌的脉冲DC 成纤维细胞和3）比较TIL与 在目标1和2中接受治疗的患者的LN。