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γ基因CDNA转移到A/Jax来源的小鼠神经母细胞瘤细胞系C1300。获得了两个感染亚克隆c - γ -3和c - γ -22分别作为低和高的生产者。这些亚克隆在体外生长模式、形态和神经丝抗原表达方面与原克隆相似，除了主要组织相容性复合体（MHC） I类抗原的表达，在表面表达水平和转录水平上都有极大的增强，而ifn - γ的产生量不同。体内致瘤性在高生成物c - γ -22中降低，但在低生成物c - γ -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率均受到抑制。因此，我们认为抑制肿瘤形成更密切地与组成型IFN-gam的产生有关，而不是与MHC抗原的表达有关，肿瘤生长速度受表面抗原高表达的影响。下一步，编码小鼠ifn - γ的cDNA被转移到一个特定的细胞毒性T淋巴细胞（CTL）克隆中，命名为E-4，用于对抗203-胶质瘤（一种源自C57/BL小鼠的20-甲基胆碱蒽诱导的胶质瘤系）。证实了外源基因产生ifn - γ对肿瘤靶向增强的作用。在5个基因转移亚克隆中，两个基因转移亚克隆的ifn - γ含量比亲本E-4高8至10倍。相应的，这两个亚克隆对203胶质瘤的杀伤活性比亲本系高2 ~ 3倍。据认为，在组成型ifn - γ产生的ctl附近的肿瘤细胞可能受到刺激，诱导或增强表面抗原的表达，包括MHC抗原以及与免疫识别相关的肿瘤相关抗原。总之，这表明逆转录病毒介导的细胞因子基因转移将有助于改进癌症的免疫治疗。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