Armstrong C A, Botella R, Galloway T H, Murray N, Kramp J M, Song I S, Ansel J C
Dermatology Service, Veterans Affairs Medical Center, Portland, Oregon, USA.
Cancer Res. 1996 May 1;56(9):2191-8.
The use of immunomodulating gene therapy in the treatment of malignant disease is under intensive investigation. In this study, we examined the potential of melanoma-derived granulocyte-macrophage colony-stimulating factor (GM-CSF) to inhibit melanoma progression in a murine model. The HGH18 murine melanoma cell line was transfected with the murine GM-CSF gene in a SV40 expression vector that resulted in melanoma clones that produced varying amounts of GM-CSF. Syngeneic mice inoculated s.c. with HFH18 parental melanoma cells or HFH18 cells transfected with the GM-CSF gene n the noncoding 3'-5' orientation [HFH18/GM-CSF(-) cells] develop large tumors that reach a mean tumor volume of 3300 mm3 by day 30. In contrast, animals inoculated with two melanoma clones producing high levels of GM-CSF [HFH18/GM-CSF(++) and HFH18/GM-CSF(+ + +)] either completely reject the tumor cells or develop tumors with a mean volume of only 40 mm3. In comparison, animals inoculated with a melanoma clone producing low levels of GM-CSF [HFH18/GM-CSF(+)] develop large tumors averaging 2000 mm3, thus demonstrating a dose-response effect of tumor inhibition by melanoma-derived GM-CSF. Additionally, vaccination with irradiated GM-CSF-producing melanoma cells conferred optimal immunogenicity against a subsequent challenge with HFH18 cells. Tissue sections from excised GM-CSF-producing tumor cell inoculation sites but not from HFH18 parental or HFH18/GM-CSF(-) inoculation sites demonstrate a dense inflammatory infiltrate composed of neutrophils, tissue macrophages, and numerous CD4- and CD8-positive lymphocytes but few melanoma cells. Large numbers of dendritic cells and cells expressing the B7-2 costimulatory molecule are detected only within HFH18/GM-CSF(+ + +) melanoma inoculation sites. Our results lend further support to clinical trials of GM-CSF gene therapy in the treatment of advanced malignant melanoma, possibly by the recruitment of dendritic antigen-presenting cells.
免疫调节基因疗法在恶性疾病治疗中的应用正在深入研究中。在本研究中，我们在小鼠模型中检测了黑色素瘤来源的粒细胞-巨噬细胞集落刺激因子（GM-CSF）抑制黑色素瘤进展的潜力。HGH18小鼠黑色素瘤细胞系用SV40表达载体中的小鼠GM-CSF基因进行转染，产生了产生不同量GM-CSF的黑色素瘤克隆。同基因小鼠皮下接种HFH18亲本黑色素瘤细胞或用非编码3'-5'方向的GM-CSF基因转染的HFH18细胞[HFH18/GM-CSF(-)细胞]，会形成大肿瘤，到第30天时平均肿瘤体积达到3300立方毫米。相比之下，接种了两个产生高水平GM-CSF的黑色素瘤克隆[HFH18/GM-CSF(++)和HFH18/GM-CSF(+++)]的动物要么完全排斥肿瘤细胞，要么形成平均体积仅为40立方毫米的肿瘤。相比之下，接种了产生低水平GM-CSF的黑色素瘤克隆[HFH18/GM-CSF(+)]的动物会形成平均体积为2000立方毫米的大肿瘤，从而证明了黑色素瘤来源的GM-CSF对肿瘤抑制的剂量反应效应。此外，用经辐射的产生GM-CSF的黑色素瘤细胞进行疫苗接种可赋予针对随后HFH18细胞攻击的最佳免疫原性。来自切除的产生GM-CSF的肿瘤细胞接种部位的组织切片，但不是来自HFH18亲本或HFH18/GM-CSF(-)接种部位的组织切片，显示出由中性粒细胞、组织巨噬细胞和大量CD4和CD8阳性淋巴细胞组成的密集炎性浸润，但黑色素瘤细胞很少。仅在HFH18/GM-CSF(+++)黑色素瘤接种部位检测到大量树突状细胞和表达B7-2共刺激分子的细胞。我们的结果进一步支持了GM-CSF基因疗法治疗晚期恶性黑色素瘤的临床试验，可能是通过募集树突状抗原呈递细胞来实现的。
