Bonnekoh B, Greenhalgh D A, Chen S H, Block A, Rich S S, Krieg T, Woo S L, Roop D R
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
J Invest Dermatol. 1998 Jun;110(6):867-71. doi: 10.1046/j.1523-1747.1998.00221.x.
The efficacy of adenovirus-mediated gene therapy for treatment of metastatic B16 melanomas, established in syngeneic C57BL/6 mice, was assessed via an ex vivo cytokine vaccine approach or via an in vivo strategy utilizing combination cytokine/herpes simplex virus-thymidine kinase (HSV-tk) suicide gene delivery and treatment with ganciclovir (GCV). In the ex vivo tumor vaccine approach, B16 melanoma cells, transduced in vitro by adenovirus containing either interleukin (IL)-2, granulocyte-macrophage colony stimulating factor (GM-CSF), or tumor necrosis factor-alpha cytokine genes and gamma irradiated, were subcutaneously injected into the flank and a distant subcutaneous challenge injection of unmodified B16 melanoma cells was performed 15 d later. Significant reductions in challenge tumor volume were observed in the IL-2 group (75% reduction; p = 0.02) and in the GM-CSF group (88% reduction; p = 0.0006), whereas the effect for tumor necrosis factor-alpha was not statistically significant. In the in vivo treatment of established melanomas, this cytokine approach was combined with a suicide gene therapy and subcutaneous B16 melanomas were directly injected with (i) IL-2/recombinant, replication-deficient adenovirus (adv) and thymidine kinase (tk)/adv, (ii) GM-CSF/adv, IL-2/adv, and tk/adv, or (iii) control beta-galactosidase (beta-gal)/adv and tk/adv. After intraperitoneal application of GCV (10 mg per kg) for 6 d, the residual tumor masses were excised and the animals challenged with unmodified B16 cells. Challenge tumor growth was reduced by 56% for the IL-2/tk/adv/GCV treatment (p = 0.041) and by 77% for the GM-CSF/IL-2/tk/adv/GCV treatment p (p = 0.037), in comparison with the beta-gal/tk/GCV control group. These data may hold significant promise for the development of effective ex vivo and in vivo gene therapy modalities to counter the highly metastatic nature of human melanoma.
通过体外细胞因子疫苗方法或体内策略，利用细胞因子/单纯疱疹病毒胸苷激酶（HSV-tk）自杀基因联合递送并使用更昔洛韦（GCV）进行治疗，评估腺病毒介导的基因疗法对同基因C57BL/6小鼠中建立的转移性B16黑色素瘤的疗效。在体外肿瘤疫苗方法中，将含有白细胞介素（IL）-2、粒细胞-巨噬细胞集落刺激因子（GM-CSF）或肿瘤坏死因子-α细胞因子基因的腺病毒在体外转导并经γ射线照射的B16黑色素瘤细胞皮下注射到侧腹，15天后进行未修饰的B16黑色素瘤细胞的远处皮下攻击注射。在IL-2组（减少75%；p = 0.02）和GM-CSF组（减少88%；p = 0.0006）中观察到攻击肿瘤体积显著减小，而肿瘤坏死因子-α的效果无统计学意义。在对已建立的黑色素瘤进行体内治疗时，这种细胞因子方法与自杀基因疗法相结合，将皮下B16黑色素瘤直接注射（i）IL-2/重组、复制缺陷型腺病毒（adv）和胸苷激酶（tk）/adv，（ii）GM-CSF/adv、IL-2/adv和tk/adv，或（iii）对照β-半乳糖苷酶（β-gal）/adv和tk/adv。腹腔内应用GCV（每千克10毫克）6天后，切除残留肿瘤块，并用未修饰的B16细胞攻击动物。与β-gal/tk/GCV对照组相比，IL-2/tk/adv/GCV治疗使攻击肿瘤生长减少56%（p = 0.041），GM-CSF/IL-2/tk/adv/GCV治疗使攻击肿瘤生长减少77%（p = 0.037）。这些数据对于开发有效的体外和体内基因治疗方式以对抗人类黑色素瘤的高转移性可能具有重要前景。
