Adoptive Immunotherapy for Malignant Brain Tumors Using Killer T Lymphocytes Enhanced by Interferon Gamma Gene Transfer

用含有干扰素-γ基因的嵌合逆转录病毒，将小鼠干扰素-γCDNA导入A/JAX来源的小鼠神经母细胞瘤细胞系C1300。获得了两个受感染的亚克隆C-Gamma-3和C-Gamma-22，分别作为低产量和高产量。这些亚克隆在神经丝的体外生长模式、形态和抗原表达方面与原始克隆相似，但主要的组相性复合体(MHC)I类抗原的表达在表面表达水平和转录水平上都显著增强，无论它们产生的干扰素-γ的量如何不同。体内致瘤性在高产量的C-伽马-22中降低，但在低产量的C-伽马-3中没有降低。与亲本系相比，两个亚克隆的体内肿瘤生长速度都受到抑制。因此，我们认为抑制肿瘤的形成与组成型干扰素-gam…更密切相关。作为下一步，编码小鼠干扰素-γ的cDNA被转移到一个特异性的细胞毒性T淋巴细胞(CTL)克隆中，命名为E-4，以对抗203-胶质瘤(C57/BL小鼠来源的20-甲基胆蒽诱导的胶质瘤系)。证实外源基因产生的干扰素-γ具有增强肿瘤靶向性的作用。在五个亚克隆中，有两个基因转移的亚克隆产生的干扰素-γ比亲本E-4高8到10倍。相应地，这两个亚克隆对203-胶质瘤的杀伤活性是亲本系的2~3倍。有人认为，在干扰素-γ产生的CTL附近的肿瘤细胞可以被刺激以诱导或增强表面抗原的表达，包括MHC抗原和与免疫识别相关的肿瘤相关抗原。综上所述，逆转录病毒介导的细胞因子基因转移有望用于改良的癌症免疫治疗。较少

A mouse ihterfebon (IFN) gamma CDNA was transferred to mouse neuroblastoma cell line, C1300 of A/Jax origin, with a chimeric retrovirus containing the IFN gamma gene. Two infected subclones C-gamma-3 and C-gamma-22 were obtained as a low and a high producers, respectively. These subclones were similar to the original clone in respects of in vitro growth pattern, morphology, and antigen expression of neurofilaments, except the expression of major hisiocompatibility complex (MHC) class I antigens, which were extremely augmented at the surface expression level as well as at the transcription level, regardless the difference in amount of their IFN-gamma production. The in vivo tumorigenicity was reduced in the high producer, C-gamma-22, but not in the low producer, C-gamma-3. The in vivo tumor growth rate was suppressed in both subclones, as compared to the parental line. It was therefore suggested that the suppression of tumor formation is more closely associated with constitutive IFN-gam … More ma production rather than the MHC antigen expression, and that the tumor growth rate is affected by the high expression of the surface antigens.As a next step, the cDNA encoding mouse IFN-gamma was transferred into a specific cytotoxic T lymphocyte (CTL) clone, designated E-4, against 203-glioma (a 20- methylcholanthrene-induced glioma line of C57/BL mouse origin). The efficacy of IFN-gamma production from the exogenous gene on augumeniation of tumor targeting was confirmed. Out of five, two gene-transferred subclones constitutively produced 8 to 10 times higher amount of IFN-gamma as compared with the parental E-4. Correspondingly, these two subclones exhibited 2 to 3 times higher killing activity against 203-glioma as compared with the parental line. It was thought that tumor cells, in the vicinity of the constitutively IFN-gamma-producing CTLs, may be stimulated to induce or enhance the expression of surface antigens, including the MHC antigens as well as the tumor associated antigens relevant to immune recognition.In summary, it was hopefully suggested that retrovirus-mediated transfer of cytokine genes would be useful for a modified immunotherapy of cancer. Less