Estasblishment of adoptive immunotherapy for cancer using T lymphocytes engneered by interleukin-2 gene

利用白细胞介素2基因工程T淋巴细胞建立癌症过继免疫疗法

基本信息

批准号：
07671321
负责人：
YAMAUE Hiroki
金额：
$ 1.6万
依托单位：
WAKAYAMA MEDICAL SCHOOL
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

项目摘要

In this study, we transduced melanoma tumor-infiltrating lymphocytes (TILs) with the IL-2 gene and darified functional characteristics of the TIL transductants. TILs transduced with 3'-end-truncated IL-2 gene (480bp) produced high amounts of IL-2 detected in supematants when compared to TILs transduced with the naive IL-2 gene containing 3'-end adenine-thymidine (AI)-rich sequences (650bp). The level of IL-2 in supematants was higher with the addition of anti-Tac antibody (Ab) to block the consumption of IL-2 by die TILs. These TILs could proliferate autonomously in the absence of exogenous IL-2, and the proliferation of TILs could be completely blocked by anti-IL-2 Ab or anti-IL-2 receptor Ab. Thus TILs transduced with IL-2 gene can proliferate through the autocrine loop. However, the expression of IL-2 from TILs transduced with the IL-2 gene was downregulated after 2 to 3 weeks of G418 selection. Our study indicates the feasibility of transduction and expression of a truncated 480-bp IL-2 gene into TILs and possiblhty of employing adoptive immunotherapy protocols using TILs modified with this IL-2 gene. Next, in a murine subcutaneus tumor model, we inoculated IL-2 gene-modified fibroblasts and/or tumor cells into syngeneic mice, and observed die tumor growth. The tumor growth was significantly inhibited in mice inoculated with parental tumor cells plus IL-2 gene-modified fibroblasts, compared with that in parental tumor cells alone. Moreover, we inoculated tumortissue to murine cecum orthotopically, and treated with IL-2 gene-modified fibroblasts plus tumor cells. The tumor volume in cecum was also significantly decreased. These results suggest that gene therapy using IL-2 gene-modified fibroblasts may be a promising strategy for a clinical application.

在这项研究中，我们用IL-2基因转导了黑色素瘤肿瘤浸润淋巴细胞（TILS），并具有TIL转导剂的darifiend型功能特征。与用含有3'-末端腺嘌呤 - 胸他胺（AI） - 幼稚的IL-2基因相比，用3'-末端截断的IL-2基因（480bp）转导的TILS在超级药物中产生了高量的IL-2。丰富的序列（650bp）。添加抗TAC抗体（AB）以阻止Die TILS的IL-2消耗，超级抗体中的IL-2水平较高。在没有外源IL-2的情况下，这些TIL可以自主扩散，并且TIL的增殖可以被抗IL-2 AB或抗IL-2受体AB完全阻塞。因此，用IL-2基因转导的TILS可以通过自分泌循环增殖。然而，在选择G418 2至3周后，用IL-2基因转导的TILS的IL-2表达下调。我们的研究表明，使用该IL-2基因修饰的TIL使用TIL进行转导480 bp IL-2基因转导和表达的可行性，并可能采用采用免疫疗法方案。接下来，在鼠皮下肿瘤模型中，我们将IL-2基因修饰的成纤维细胞和/或肿瘤细胞接种到合成小鼠中，并观察到死亡的肿瘤生长。与单独的亲本肿瘤细胞相比，与亲本肿瘤细胞和IL-2基因改性成纤维细胞接种的小鼠相比，肿瘤生长受到显着抑制。此外，我们将肿瘤接种到原始的鼠盲鼠，并用IL-2基因修饰的成纤维细胞和肿瘤细胞进行治疗。盲肠中的肿瘤体积也显着减少。这些结果表明，使用IL-2基因改性成纤维细胞的基因治疗可能是临床应用的有前途的策略。