Fernandez Marilyn, Porosnicu Mercedes, Markovic Dubravka, Barber Glen N
Department of Microbiology and Immunology and Sylvester Comprehensive Cancer Center, University of Miami School of Medicine, Miami, Florida 33136, USA.
J Virol. 2002 Jan;76(2):895-904. doi: 10.1128/jvi.76.2.895-904.2002.
We report here the generation of recombinant vesicular stomatitis virus (VSV) able to produce the suicide gene product thymidine kinase (TK) or cytokine interleukin 4 (IL-4). In vitro cells infected with the engineered viruses expressed remarkably high levels of biologically active TK or IL-4 and showed no defects in replication compared to the wild-type virus. Recombinant viruses retained their ability to induce potent apoptosis in a variety of cancer cells, while normal cells were evidently more resistant to infection and were completely protected by interferon. Significantly, following direct intratumoral inoculation, VSV expressing either TK or IL-4 exhibited considerably more oncolytic activity against syngeneic breast or melanoma tumors in murine models than did the wild-type virus or control recombinant viruses expressing green fluorescent protein (GFP). Complete regression of a number of tumors was achieved, and increased granulocyte-infiltrating activity with concomitant, antitumor cytotoxic T-cell responses was observed. Aside from discovering greater oncolytic activity following direct intratumoral inoculation, however, we also established that VSV expressing IL-4 or TK, but not GFP, was able to exert enhanced antitumor activity against metastatic disease. Following intravenous administration of the recombinant viruses, immunocompetent BALB/c mice inoculated with mammary adenocarcinoma exhibited prolonged survival against lethal lung metastasis. Our data demonstrate the validity of developing novel types of engineered VSV for recombinant protein production and as a gene therapy vector for the treatment of malignant and other disease.
我们在此报告能够产生自杀基因产物胸苷激酶（TK）或细胞因子白细胞介素4（IL-4）的重组水疱性口炎病毒（VSV）的构建。与野生型病毒相比，用工程病毒感染的体外细胞显著高水平表达生物活性的TK或IL-4，并且在复制方面没有缺陷。重组病毒保留了在多种癌细胞中诱导有效凋亡的能力，而正常细胞显然对感染更具抗性，并受到干扰素的完全保护。值得注意的是，在直接瘤内接种后，表达TK或IL-4的VSV在小鼠模型中对同基因乳腺或黑色素瘤肿瘤的溶瘤活性比野生型病毒或表达绿色荧光蛋白（GFP）的对照重组病毒显著更高。实现了许多肿瘤的完全消退，并观察到粒细胞浸润活性增加以及伴随的抗肿瘤细胞毒性T细胞反应。然而，除了发现直接瘤内接种后具有更高的溶瘤活性外，我们还证实表达IL-4或TK而非GFP的VSV能够对转移性疾病发挥增强的抗肿瘤活性。在静脉内给予重组病毒后，接种乳腺腺癌的免疫活性BALB/c小鼠对致命性肺转移表现出延长的生存期。我们的数据证明了开发新型工程化VSV用于重组蛋白生产以及作为治疗恶性疾病和其他疾病的基因治疗载体的有效性。
