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

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AdC MVLacZ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）、（1 86.91±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。

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

Objective To investigate the anti-tumor effects of combined recombinant murine interleukin-12 and GM-CSF gene adenovirus vector(AdCMVIL-12,AdCMVGMCSF) 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 GMCSF 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-CSF group (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.