IMMUNE RESPONSES INDUCED BY GENE TRANSFER

基因转移引起的免疫反应

基本信息

批准号：
6237409
负责人：
ALFRED E CHANG
金额：
$ 28.84万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-06-01 至 1998-03-31
项目状态：
已结题

项目摘要

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.

根据我们对上一个资助期的研究，我们已经能够开发两种上调免疫的遗传方法对肿瘤的反应性。第一种方法涉及体内基因将同种异体 MHC 基因转移到动物和动物体内的肿瘤中人类研究发现可以可靠地导致转基因表达。更重要的是，我们能够增强抗肿瘤反应性肿瘤内和引流淋巴结 (LN) 内；并观察到肿瘤特定患者的回归。第二种方法涉及使用经基因改造可分泌 GM-CSF 的自体肿瘤疫苗增强引流淋巴结中局部免疫反应的方法。我们目前正在进行一项第一阶段研究，以评估转基因生物的使用 CSF 转导肿瘤作为疫苗，产生免疫 LN 细胞过继性免疫疗法。该项目的主要重点是开发肿瘤反应性 T 细胞可以在体外扩展用于临床治疗。我们建议评估源自 TIL 的体外和体内肿瘤反应性病灶内 HLA-B7 基因治疗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 限制诱导的细胞反应基因转移。该项目的另一个方面将是树突状细胞（DC）的应用，其将被用作肿瘤疫苗的成分使 LN 细胞敏感，以进行后续治疗过继性免疫疗法。 DC拥有独特的素养能力用于体外和体内免疫反应的 T 细胞。 DC研究从皮肤肿瘤中分离出来的物质已证明其功能能力可被免疫抑制细胞因子抑制。这有 GM-CSF 的存在逆转了这两种情况 B7-1 和 B7-2 共刺激分子。我们计划评估用肿瘤脉冲 DC 体内引发的 LN 细胞的抗肿瘤反应性与黑色素瘤患者的 DM-CSF 成纤维细胞混合结直肠癌。该项目的具体目标是： 1) 评估源自直接修饰的肿瘤的 TIL 的免疫反应性利用同种异体 I 类 MHC 基因进行体内基因转移，2) 评估体内 T 细胞的免疫反应性混合或不混合 GM-CSF 分泌的脉冲 DC 成纤维细胞，以及 3) 比较 TIL 与在目标 1 和 2 中接受治疗的患者中启动 LN。