Helper virus-free herpes simplex virus type 1 amplicon vectors for granulocyte-macrophage colony-stimulating factor-enhanced vaccination therapy for experimental glioma.
Helper virus-free herpes simplex virus type 1 amplicon vectors for granulocyte-macrophage colony-stimulating factor-enhanced vaccination therapy for experimental glioma.
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DOI：
10.1089/10430340050057503
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发表时间：
2000-07
影响因子：
4.2
通讯作者：
Ulrich Herrlinger;A. Jacobs;Ariel Quiñones;C. Woiciechowsky;Miguel Sena-Esteves;N. Rainov;C. Fraefel;X. Breakefield
中科院分区：
文献类型：
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作者：
Ulrich Herrlinger;A. Jacobs;Ariel Quiñones;C. Woiciechowsky;Miguel Sena-Esteves;N. Rainov;C. Fraefel;X. Breakefield
胶质瘤细胞，这是逆转录病毒转导分泌粒细胞-巨噬细胞集落刺激因子（GM-CSF），皮下接种治疗，先前已证明有效的C57 BL/6小鼠窝藏脑内GL 261胶质瘤。然而，人类神经胶质瘤的临床离体基因治疗将是困难的，因为通过逆转录病毒载体的转基因递送仅发生在分裂细胞中，并且离体神经胶质瘤细胞具有低生长分数。为了避免这个问题，使用无辅助病毒的单纯疱疹病毒1型（HSV-1）扩增子载体。当用HSV-1扩增子载体以MOI为1感染人胶质母细胞瘤细胞的原代培养物时，超过90%的分裂和非分裂细胞被转导。当细胞在Polybrene存在下用携带GM-CSF cDNA的扩增子载体HSVGM感染时，在感染后的前24小时内GM-CSF分泌到培养基中为1026 ng/10（6）个细胞，而模拟感染的细胞不分泌可检测的GM-CSF。在脑内植入10（6）个野生型GL 261细胞之前7天，用5 × 10（5）个经辐照的HSVGM-转导的GL 261细胞皮下接种C57 BL/6小鼠产生60%的长期存活（>80天），类似于通过用逆转录病毒GM-CSF-转导的GL 261细胞接种获得的50%的长期存活。相比之下，用相同数量的非转导的GL 261细胞或用无辅助病毒包装的HSV-1扩增子载体感染的GL 261细胞接种的动物仅显示10%的长期存活。总之，无辅助病毒的HSV-1扩增子载体似乎是有效的，用于神经胶质瘤的尼古丁增强的疫苗接种治疗，具有的优点是分裂和非分裂肿瘤细胞都可以被感染，没有病毒蛋白表达，并且这些载体是安全的，与临床使用相容。
Subcutaneous vaccination therapy with glioma cells, which are retrovirally transduced to secrete granulocyte-macrophage colony-stimulating factor (GM-CSF), has previously proven effective in C57BL/6 mice harboring intracerebral GL261 gliomas. However, clinical ex vivo gene therapy for human gliomas would be difficult, as transgene delivery via retroviral vectors occurs only in dividing cells and ex vivo glioma cells have a low growth fraction. To circumvent this problem, a helper virus-free herpes simplex virus type 1 (HSV-1) amplicon vector was used. When primary cultures of human glioblastoma cells were infected with HSV-1 amplicon vectors at an MOI of 1, more than 90% of both dividing and nondividing cells were transduced. When cells were infected with an amplicon vector, HSVGM, bearing the GM-CSF cDNA in the presence of Polybrene, GM-CSF secretion into the medium during the first 24 hr after infection was 1026 ng/10(6) cells, whereas mock-infected cells did not secrete detectable GM-CSF. Subcutaneous vaccination of C57BL/6 mice with 5 x 10(5) irradiated HSVGM-transduced GL261 cells 7 days prior to intracerebral implantation of 10(6) wild-type GL261 cells yielded 60% long-term survivors (>80 days), similar to the 50% long-term survivors obtained by vaccination with retrovirally GM-CSF-transduced GL261 cells. In contrast, animals vaccinated with the same number of nontranduced GL261 cells or with GL261 cells infected with helper virus-free packaged HSV-1 amplicon vectors carrying no transgene showed only 10% long-term survivors. In conclusion, helper virus-free HSV-1 amplicon vectors appear to be effective for cytokine-enhanced vaccination therapy of glioma, with the advantages that both dividing and nondividing tumor cells can be infected, no viral proteins are expressed, and these vectors are safe and compatible with clinical use.