Inactivation of the UL37 Deamidase Enhances Virus Replication and Spread of the HSV-1(VC2) Oncolytic Vaccine Strain and Secretion of GM-CSF.
Inactivation of the UL37 Deamidase Enhances Virus Replication and Spread of the HSV-1(VC2) Oncolytic Vaccine Strain and Secretion of GM-CSF.
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DOI：
10.3390/v15020367
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发表时间：
2023-01-27
期刊：
影响因子：
--
通讯作者：
Kousoulas KG
中科院分区：
文献类型：
--
作者：
Clark CM;Jambunathan N;Collantes TMA;Kousoulas KG
在糖蛋白K （gK）和膜蛋白UL20的氨基末端进行特异性缺失，使病毒无法进入神经元并建立潜伏期，从而设计了HSV-1 （VC2）减毒活疫苗株。VC2在上皮细胞培养中有效复制，但与亲本HSV-1 (F)野生型病毒相比，其病毒滴度较低，病毒斑块较小。VC2是在小鼠和豚鼠中针对HSV-1和HSV-2感染的有效减毒活疫苗，以及在小鼠模型中针对黑色素瘤和乳腺癌的抗肿瘤免疫治疗和溶瘤病毒。在此之前，我们报道了gK/UL20复合物与UL37被膜蛋白相互作用，这种相互作用对于病毒粒子的细胞内包膜和出口至关重要。为了研究UL37脱酰胺酶功能的潜在作用，在HSV-1(F)和HSV-1（VC2）遗传背景下，用C819S替代构建重组病毒FC819S和VC2C819S，分别灭活UL37预测的脱酰胺酶活性位点。FC819S与HSV-1(F)复制到相似的水平，并产生相似大小的病毒斑块。相比之下，VC2C819S复制增强，病毒斑块大小增加，接近野生型HSV-1(F)病毒。与HSV-1(F)相比，细胞培养物感染FC819S导致多种细胞系（包括HEp2细胞和癌细胞系、DU145（前列腺）和Panc 04.03（胰腺））以及小鼠原代腹膜细胞中GM-CSF分泌增强。这些细胞系感染VC2引起的GM-CSF分泌水平与感染FC819S相似。然而，与VC2病毒相比，VC2C819S病毒没有进一步增强GM-CSF的分泌。这些结果表明，UL37脱酰胺与gK/UL20复合物一起发挥作用，促进病毒复制和GM-CSF分泌。
The HSV-1 (VC2) live-attenuated vaccine strain was engineered with specific deletions in the amino termini of glycoprotein K (gK) and membrane protein UL20, rendering the virus unable to enter neurons and establish latency. VC2 replicates efficiently in epithelial cell culture but produces lower viral titers and smaller viral plaques than its parental HSV-1 (F) wild-type virus. VC2 is an effective live-attenuated vaccine against HSV-1 and HSV-2 infections in mice and guinea pigs and an anti-tumor immunotherapeutic and oncolytic virus against melanoma and breast cancer in mouse models. Previously, we reported that the gK/UL20 complex interacts with the UL37 tegument protein, and this interaction is essential for virion intracellular envelopment and egress. To investigate the potential role of the UL37 deamidase functions, the recombinant virus FC819S and VC2C819S were constructed with a C819S substitution to inactivate the UL37 predicted deamidase active site on an HSV-1(F) and HSV-1(VC2) genetic background, respectively. FC819S replicated to similar levels with HSV-1(F) and produced similar size viral plaques. In contrast, VC2C819S replication was enhanced, and viral plaques increased in size, approaching those of the wild-type HSV-1(F) virus. FC819S infection of cell cultures caused enhanced GM-CSF secretion in comparison to HSV-1(F) across several cell lines, including HEp2 cells and cancer cell lines, DU145 (prostate) and Panc 04.03 (pancreas), and primary mouse peritoneal cells. VC2 infection of these cell lines caused GM-CSF secretion at similar levels to FC819S infection. However, the VC2C819S virus did not exhibit any further enhancement of GM-CSF secretion compared to the VC2 virus. These results suggest that the UL37 deamidation functions in conjunction with the gK/UL20 complex to facilitate virus replication and GM-CSF secretion.
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DOI：
10.1111/tra.12408
发表时间：
2016-09
期刊：
Traffic (Copenhagen, Denmark)
影响因子：
--
作者：
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通讯作者：
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影响因子：
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DOI：
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发表时间：
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影响因子：
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