Nishihara K, Barth R F, Wilkie N, Lang J C, Oda Y, Kikuchi H, Everson M P, Lotze M T
Department of Pathology, Ohio State University, Columbus 43210, USA.
Cancer Gene Ther. 1995 Jun;2(2):113-24.
Genetically engineered monocytes and macrophages may have potential as effector cells for the adoptive immunotherapy of cancer. As a first step, we have transfected the genes encoding either mouse interferon (IFN)-gamma, human interleukin (IL)-6, mouse IL-4, or mouse tumor necrosis factor (TNF)-alpha into the mouse macrophage cell line, J774A.1 cells using retroviral vectors. In vitro activation of J774A.1 cells by gene modification was assessed by morphological changes, proliferative activity was determined by [3H]-TdR uptake, and cytolytic activity was assessed using an 18-hour chromium-51 (51Cr) release assay. In vivo tumoricidal activity was studied by means of local adoptive immunotherapy using intratumoral injection of transfected effector cells. IFN-gamma gene-transfected J774A.1 [J7(IFN-gamma)] cells developed filamentous processes, increased doubling times, and enhanced tumoricidal activity against three tumor cell lines: the TNF-sensitive fibrosarcoma line WEHI 164 and the TNF-alpha-resistant cell lines B16 melanoma and C1300 neuroblastoma. IL-6-, TNF-alpha-, and IL-4-gene-transfected J774A.1 cells also had augmented tumoricidal activity but did not display any changes in morphology or growth. Cytolytic activity was markedly reduced after the addition of anti-TNF-alpha antibodies. Cytolytic J7(IFN-gamma) cells showed upregulated expression of TNF-alpha messenger RNA. After intratumoral injection of J7(IL-4) and J7(IFN-gamma) cell mixtures, 50% of established B16 melanomas were rejected by C57BL/6 mice, thereby demonstrating synergistic killing. Further studies on gene-transfected macrophages should better define their potential usefulness in tumor immunotherapy.
基因工程改造的单核细胞和巨噬细胞可能具有作为癌症过继性免疫治疗效应细胞的潜力。作为第一步，我们使用逆转录病毒载体将编码小鼠干扰素（IFN）-γ、人白细胞介素（IL）-6、小鼠IL-4或小鼠肿瘤坏死因子（TNF）-α的基因转染到小鼠巨噬细胞系J774A.1细胞中。通过形态学变化评估基因修饰对J774A.1细胞的体外激活，通过[3H]-TdR摄取测定增殖活性，并使用18小时的铬-51（51Cr）释放试验评估细胞溶解活性。通过瘤内注射转染的效应细胞进行局部过继性免疫治疗来研究体内杀瘤活性。IFN-γ基因转染的J774A.1 [J7(IFN-γ)]细胞形成丝状突起，倍增时间增加，并增强了对三种肿瘤细胞系的杀瘤活性：TNF敏感的纤维肉瘤系WEHI 164以及TNF-α抗性细胞系B16黑色素瘤和C1300神经母细胞瘤。IL-6、TNF-α和IL-4基因转染的J774A.1细胞也具有增强的杀瘤活性，但在形态或生长方面未显示任何变化。加入抗TNF-α抗体后，细胞溶解活性明显降低。具有细胞溶解活性的J7(IFN-γ)细胞显示TNF-α信使RNA的表达上调。在瘤内注射J7(IL-4)和J7(IFN-γ)细胞混合物后，50%已建立的B16黑色素瘤被C57BL/6小鼠排斥，从而证明了协同杀伤作用。对基因转染巨噬细胞的进一步研究应能更好地确定它们在肿瘤免疫治疗中的潜在用途。
