文章：

用GM-CSF基因武装溶瘤呼肠孤病毒增强免疫

Arming oncolytic reovirus with GM-CSF gene to enhance immunity 

原文发布日期：2018-11-23 

英文摘要：

Oncolytic reovirus administration has been well tolerated by cancer patients in clinical trials. However, its anti-cancer efficacy as a monotherapy remains to be augmented. We and others have previously demonstrated the feasibility of producing replication-competent reoviruses expressing a heterologous transgene. Here, we describe the production of recombinant reoviruses expressing murine (mm) or human (hs) GM-CSF (rS1-mmGMCSF and rS1-hsGMCSF, respectively). The viruses could be propagated up to 10 passages while deletion mutants occurred only occasionally. In infected cell cultures, the secretion of GM-CSF protein (up to 481 ng/106 cells per day) was demonstrated by ELISA. The secreted mmGM-CSF protein was functional in cell culture, as demonstrated by the capacity to stimulate the survival and proliferation of the GM-CSF-dependent dendritic cell (DC) line D1, and by its ability to generate DCs from murine bone marrow cells. Importantly, in a murine model of pancreatic cancer we found a systemic increase in DC and T-cell activation upon intratumoral administration of rS1-mmGMCSF. These data demonstrate that reoviruses expressing functional GM-CSF can be generated and have the potential to enhance anti-tumor immune responses. The GM-CSF reoviruses represent a promising new agent for use in oncolytic virotherapy strategies. 

摘要翻译： 

在临床试验中，溶瘤呼肠孤病毒对癌症患者表现出良好的耐受性。然而其作为单一疗法的抗肿瘤效力仍有待提升。我们与其他研究团队此前已证实，生产能表达异源转基因且具备复制能力的呼肠孤病毒具有可行性。本研究成功制备出表达小鼠（mm）或人源（hs）GM-CSF的重组呼肠孤病毒（分别为rS1-mmGMCSF和rS1-hsGMCSF）。这些病毒可连续传代培养至10代，且仅偶尔出现缺失突变。通过ELISA检测发现，在感染细胞培养物中GM-CSF蛋白分泌量可达每日481纳克/106个细胞。分泌的小鼠GM-CSF蛋白在细胞培养中具有功能活性，表现为能刺激GM-CSF依赖性树突状细胞（DC）系D1的存活与增殖，并能诱导小鼠骨髓细胞分化为树突状细胞。重要的是，在胰腺癌小鼠模型中，瘤内注射rS1-mmGMCSF可引起树突状细胞和T细胞活化的系统性增强。这些数据表明，能够表达功能性GM-CSF的呼肠孤病毒不仅可被成功制备，还具有增强抗肿瘤免疫反应的潜力。GM-CSF呼肠孤病毒为溶瘤病毒治疗策略提供了一种前景广阔的新型制剂。

原文链接：

Arming oncolytic reovirus with GM-CSF gene to enhance immunity