重组鼠白细胞介素12与粒细胞-巨噬细胞集落刺激因子基因联合治疗小鼠B16黑色素瘤

Effects of recombinant murine interlukin-12 and GM-CSF gene adenovirus vectors on B16 melanoma

目的:研究重组鼠白细胞介素12(AdCMVIL-12)与粒细胞巨噬细胞集落刺激因子(AdCMVGM-CSF)的腺病毒载体联合应用对B16黑色素瘤的抗肿瘤作用。方法:将B16黑色素瘤细胞接种于C57/BL6小鼠右侧背部皮下后,待瘤体直径达5mm时将荷瘤小鼠随机分为4组,分别为AdCMVIL-12组、AdCMVGM-CSF组、AdCMVIL-12加AdCMVGM-CSF组(联合治疗组)和AdCMVLacZ组(对照组),每组12只,并于肿瘤组织局部分别多点注射相对应剂量的腺病毒载体,观察各组小鼠肿瘤的生长情况及其诱导的细胞免疫反应,检测各组小鼠血清IL-12和GM-CSF表达水平的变化。结果:病理学观察,联合治疗组肿瘤组织内可见大片凝固性坏死,有大量的淋巴细胞、巨噬细胞浸润。治疗30 d后,联合治疗组肿瘤平均体积为(60.73±11.36)mm3,与对照组、AdCMVIL-12组和AdCMVGM-CSF组[分别为(675.18±80.59)、(186.91±19.15)和(223.45±23.11)mm3]比较差异均有显著性(P<0.01)。同时,AdCMVIL-12与AdCMVGM-CSF联合基因治疗可有效延长IL-12和GM-CSF的表达时间,并且对B16黑色素瘤细胞具有很强的特异性杀伤作用,杀伤率为(69.53±9.20)%。结论:IL-12与GM-CSF联合基因治疗可增强荷B16黑色素瘤小鼠的抗肿瘤免疫反应,并能有效降低单独应用AdCMVIL-12的毒副作用。

Objective To investigate the anti-tumor effects of combined recombinant murine interleukin-12 and GM- CSF gene adenovirus vector （AdCMVIL-12, AdCMVGM-CSF） on B16 melanoma. Methods B16 melanoma cells were injected subcutaneous at right back of C57/BL6 mice, 48 tumor bearing mice were divided into 4 groups at random, when tumor diameter attained to 5 mm, AdCMVIL-12, AdCMVGM-CSF and AdCMVIL-12 ＋ AdCMVGM -CSF and AdCMVLacZ （control） were injected in different groups, tumor growth and immune responses of each group were tested, and the IL-12 and GM-CSF levels in serum were observed. Results In combined group, necrosis, lymphocytes and macrophages were found in tumor tissues. 30 d after treatment, the average volume in combined group was （60.73± 11.36） mm^3, compared with AdCMVLacZ （control） group, and AdCMVIL-12 group, AdCMVGM-CSFgroup [ （675. 18 ±80.59）, （186.91 ±19.15）, （223.45±23. 11） mm^3], there were significant differences （P〈0.01） . Combined IL-12 and GM-CSF gene therapy could enhanced anti-tumor immune response of B16 melanoma bearing mice and increased the expressing time of IL-12 and GM-CSF, and had very strong specific kill effect. Kill probability was （69.53± 9.20）%. Conclusion Combined IL-12 and GM-CSF gene therapy can enhance the anti-tumor immune response of B16 melanoma bearing mice and decrease side-effect of using AdCMVIL-12.