Production Strategies for Pentamer-Positive Subviral Dense Bodies as a Safe Human Cytomegalovirus Vaccine
Production Strategies for Pentamer-Positive Subviral Dense Bodies as a Safe Human Cytomegalovirus Vaccine
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DOI：
10.3390/vaccines7030104
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发表时间：
2019-09-01
期刊：
影响因子：
7.8
通讯作者：
Plachter, Bodo
中科院分区：
文献类型：
--
作者：
Gogesch, Patricia;Penner, Inessa;Plachter, Bodo
人巨细胞病毒（HCMV）感染与儿童产前传播和免疫抑制个体病毒再激活后的严重临床表现相关。长期以来一直要求开发HCMV疫苗，但仍然没有获得许可的产品。亚病毒致密体（DB）在临床前模型中具有免疫原性，因此是一种有希望的HCMV候选疫苗。最近，我们在实验室菌株Towne的基础上建立了一种病毒，该病毒合成了大量含有五聚体蛋白复合物gH/gL/UL128-131 （Towne- ul130rep Delta GFP）的DB。这里介绍的工作重点是提供基于该菌株生产安全疫苗的战略。建立了符合gmp的DB生产协议。此外，通过删除UL25开放阅读框，对DB产生菌株town - ul130rep进行了衰减。额外的遗传修饰旨在通过使用Shield-1/FKBP不稳定系统有条件地表达pUL51来消除其在体内复制的能力。我们进一步表明，终端酶抑制剂letermovir可用于减少传染性病毒污染的DB疫苗超过两个数量级。综上所述，本文提供的策略允许生产用于临床试验的安全且具有免疫原性的DB疫苗。
Infections with the human cytomegalovirus (HCMV) are associated with severe clinical manifestations in children following prenatal transmission and after viral reactivation in immunosuppressed individuals. The development of an HCMV vaccine has long been requested but there is still no licensed product available. Subviral dense bodies (DB) are immunogenic in pre-clinical models and are thus a promising HCMV vaccine candidate. Recently, we established a virus based on the laboratory strain Towne that synthesizes large numbers of DB containing the pentameric protein complex gH/gL/UL128-131 (Towne-UL130rep Delta GFP). The work presented here focuses on providing strategies for the production of a safe vaccine based on that strain. A GMP-compliant protocol for DB production was established. Furthermore, the DB producer strain Towne-UL130rep was attenuated by deleting the UL25 open reading frame. Additional genetic modifications aim to abrogate its capacity to replicate in vivo by conditionally expressing pUL51 using the Shield-1/FKBP destabilization system. We further show that the terminase inhibitor letermovir can be used to reduce infectious virus contamination of a DB vaccine by more than two orders of magnitude. Taken together, strategies are provided here that allow for the production of a safe and immunogenic DB vaccine for clinical testing.