瘤内/瘤周注射巨噬细胞集落刺激因子或(和)干扰素γ基因转染的巨噬细胞对肿瘤的治疗作用

Antitumor effect of M CSF and/or IFN γ gene cotransfected macrophages by intratumoral injection

目的观察巨噬细胞集落刺激因子（ＭＣＳＦ）和干扰素γ（ＩＦＮγ）基因单独或联合转染的巨噬细胞（ｍａｃｒｏｐｈａｇｅ，Ｍ）对局部黑色素瘤的治疗效果及相关免疫机理。方法荷瘤小鼠经基因转染的巨噬细胞治疗后，观察其长期存活期并检测其体内抗肿瘤免疫功能。用４小时５１Ｃｒ释放法检测荷瘤小鼠脾细胞自然杀伤细胞（ＮＫ）、细胞毒Ｔ淋巴细胞（ＣＴＬ）活性，间接ＭＴＴ法检测荷瘤小鼠腹腔巨噬细胞的杀伤活性，常规方法检测脾细胞诱导上清中肿瘤坏死因子（ＴＮＦ）、白细胞介素２（ＩＬ２）、γ干扰素（ＩＦＮγ）活性，局部肿瘤经治疗后行常规病理分析。结果经ＩＦＮγ基因及联合基因转染的巨噬细胞治疗后的荷瘤小鼠有２５％能长期存活，体外诱导的ＣＴＬ和小鼠腹腔巨噬细胞杀伤活性显著增强，脾细胞经诱导产生的细胞因子有不同程度的增加，与对照组相比差别显著。常规病理显示，ＭＣＳＦ和ＩＦＮγ联合基因转染的巨噬细胞治疗的小鼠肿瘤局部有大量的淋巴细胞浸润。结论ＭＣＳＦ和ＩＦＮγ基因转染的巨噬细胞瘤内瘤周注射对肿瘤有较好的治疗效果，其机制除了与巨噬细胞在局部直接接触杀伤瘤细胞有关外。

Objective To investigate the in vivo antitumor effects of M CSF and/or IFN γ gene transfected macrophages and their related immunological mechanisms. Methods Survival time and antitumor effects in vivo of tumor bearing mice treated with gene transfected macrophages were observed. NK cell and CTL activity was detected with 4hr 51 Cr release assay , cytotoxicity of macrophages was detected with indirect MTT methods, activity of IL 2,TNF,IFN γ were detected by routine methods. Results 25% of the tumor bearing mice after treatment with IFN γ gene transfected, M CSF and IFN γ gene co transfected macrophages survived more than 3 month. The CTL and macrophage activity of the tumor bearing mice was enhanced. Levels of TNF,IL 2,IFN γ in the supernatants of splenocytes induced in vitro were also increased. Histologic analyses of the tumor tissue after treatment with M CSF and/or IFN γ gene transfected macrophages showed a large amount of lymphocyte infiltration. Conclusion IFN γ and M CSF gene co transfection into macrophages are proved to enhance their antitumor potential efficiently through direct augmentation of their tumoricidal cytotoxicity and induction of host antitumor immunity.