Kim S H, Carew J F, Kooby D A, Shields J, Entwisle C, Patel S, Shah J P, Fong Y
Department of Otorhinolaryngology, Yonsei University College of Medicine, Seoul, Korea.
Cancer Gene Ther. 2000 Sep;7(9):1279-85. doi: 10.1038/sj.cgt.7700231.
Herpes simplex type 2-defective infectious single-cycle (DISC) viruses are attenuated viruses that were originally produced as viral vaccines; however, these viruses are also efficient gene transfer vehicles. The main goals of this study were to examine determinants of the gene transfer by using DISC virus for squamous cancer and to evaluate the antitumoral efficacy of vaccination with tumor cells modified by DISC viruses carrying a combination of immunomodulatory genes (interleukin-2 (IL-2), granulocyte-macrophage colony-stimulating factor (GM-CSF), B7-1) in a model of squamous cell cancer (SCCVII) in C3H/HeJ mice. SCCVII cells transduced by DISC viruses (multiplicity of infection of 10) carrying the IL-2 or GM-CSF gene produced nanogram quantities of IL-2 or GM-CSF per 10(6) cells. Irradiated (5,000 cGy, 10,000 cGy) cells secreted levels of GM-CSF or IL-2 that were comparable with nonirradiated cells. In vivo vaccination using tumor cells transduced ex vivo with DISC-IL2 or DISC-GMCSF resulted in protection against subsequent tumor challenge (P < .01), with DISC-GMCSF-transduced, irradiated tumor cells showing the greatest effects (P < .001). Marked growth arrest also was noted in established tumors after direct injection of DISC-GMCSF (P < .001). These data demonstrate that (a) DISC virus is capable of efficient gene transfer, (b) GM-CSF-secreting genetically modified tumor vaccine protects against tumor cell challenge and suppresses tumor growth, and (c) intratumoral injection of DISC-GMCSF significantly suppresses the growth of established tumors. These results not only confirm clinically relevant gene transfer but also demonstrate that the gene transfer is an effective anti-cancer therapy.
2型单纯疱疹病毒缺陷感染单周期（DISC）病毒是减毒病毒，最初作为病毒疫苗生产；然而，这些病毒也是高效的基因传递载体。本研究的主要目的是通过使用DISC病毒研究鳞状细胞癌基因传递的决定因素，并在C3H/HeJ小鼠的鳞状细胞癌（SCCVII）模型中评估用携带免疫调节基因组合（白细胞介素-2（IL-2）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、B7-1）的DISC病毒修饰的肿瘤细胞进行疫苗接种的抗肿瘤效果。携带IL-2或GM-CSF基因的DISC病毒（感染复数为10）转导的SCCVII细胞每10⁶个细胞产生纳克量的IL-2或GM-CSF。经辐照（5000 cGy、10000 cGy）的细胞分泌的GM-CSF或IL-2水平与未辐照细胞相当。使用经DISC-IL2或DISC-GMCSF体外转导的肿瘤细胞进行体内疫苗接种可预防随后的肿瘤攻击（P <.01），其中经DISC-GMCSF转导、辐照的肿瘤细胞效果最佳（P <.001）。直接注射DISC-GMCSF后，在已形成的肿瘤中也观察到明显的生长停滞（P <.001）。这些数据表明：（a）DISC病毒能够进行高效的基因传递；（b）分泌GM-CSF的基因修饰肿瘤疫苗可预防肿瘤细胞攻击并抑制肿瘤生长；（c）瘤内注射DISC-GMCSF可显著抑制已形成肿瘤的生长。这些结果不仅证实了临床相关的基因传递，还表明基因传递是一种有效的抗癌疗法。
