Rosenthal F M, Cronin K, Bannerji R, Golde D W, Gansbacher B
Department of Medicine, Memorial Sloan-Kettering Cancer Center, New York, NY 10021.
Blood. 1994 Mar 1;83(5):1289-98.
Therapeutic models using gene transfer into tumor cells have pursued three objectives: (1) to induce rejection of the tumor transduced with therapeutic genes, (2) to induce immune-mediated regression of metastatic disease, and (3) to induce long-lasting immunity to protect against challenge with tumor cells or clinical regrowth of micrometastatic disease. Because in vivo therapy for patients with cancer using gene transfer would, as a first step, attempt to eliminate the existing tumor, we have investigated whether antitumor immunity induced by tumor cells secreting a single cytokine could be increased by cotransfer of a second cytokine gene. To test this approach, CMS-5, a murine fibrosarcoma, was transduced with retroviral vectors carrying interleukin-2 (IL-2), interferon-gamma (IFN-gamma), or granulocyte-macrophage-colony-stimulating factor (GM-CSF) cDNA alone or IL-2 cDNA in combination with IFN-gamma or GM-CSF cDNA. Single cytokine-secreting clones were selected to match levels of cytokine production by double cytokine-secreting clones so that similar amounts of cytokine were secreted. IFN-gamma- and IL-2/IFN-gamma-secreting CMS-5 cells showed increased levels of major histocompatability complex class I expression compared with IL-2- and GM-CSF-secreting or parental CMS-5 cells, IL-2/IFN-gamma-secreting CMS-5 cells were always rejected by syngeneic mice, whereas the same number of CMS-5 cells secreting only one of these cytokines or mixtures of single cytokine-secreting CMS-5 cells were not rejected. In vivo depletion of CD4+, CD8+, or natural-killer effector cell subpopulations showed that CD8+ cytotoxic T cells were primarily responsible for rejection of IL-2/IFN-gamma-transduced tumor cells. Our data show the successful use of a single retroviral vector to stably transduce two cytokine genes into the same tumor cell, leading to an increased effect on the in vivo induction of antitumor immunity.
（1）诱导对转导了治疗性基因的肿瘤产生排斥反应；（2）诱导免疫介导的转移性疾病消退；（3）诱导持久免疫以防止肿瘤细胞攻击或微转移性疾病的临床复发。由于使用基因转移对癌症患者进行体内治疗首先会尝试消除现有的肿瘤，我们研究了通过共转移第二个细胞因子基因是否可以增强由分泌单一细胞因子的肿瘤细胞诱导的抗肿瘤免疫力。为了测试这种方法，用携带白细胞介素-2（IL-2）、干扰素-γ（IFN-γ）或粒细胞-巨噬细胞集落刺激因子（GM-CSF）cDNA的逆转录病毒载体单独转导或与IFN-γ或GM-CSF cDNA联合转导小鼠纤维肉瘤CMS-5。选择分泌单一细胞因子的克隆，使其与分泌双细胞因子的克隆的细胞因子产生水平相匹配，以便分泌相似量的细胞因子。与分泌IL-2和GM-CSF的或亲本CMS-5细胞相比，分泌IFN-γ和IL-2/IFN-γ的CMS-5细胞显示出主要组织相容性复合体I类表达水平增加，分泌IL-2/IFN-γ的CMS-5细胞总是被同基因小鼠排斥，而分泌这些细胞因子之一的相同数量的CMS-5细胞或分泌单一细胞因子的CMS-5细胞混合物则未被排斥。体内去除CD4 +、CD8 +或自然杀伤效应细胞亚群表明，CD8 +细胞毒性T细胞主要负责排斥IL-2/IFN-γ转导的肿瘤细胞。我们的数据表明成功使用单个逆转录病毒载体将两个细胞因子基因稳定转导到同一肿瘤细胞中，从而增强了体内诱导抗肿瘤免疫的效果。
