A chemical method for generating live-attenuated, replication-defective DNA viruses for vaccine development.
A chemical method for generating live-attenuated, replication-defective DNA viruses for vaccine development.
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DOI：
10.1016/j.crmeth.2022.100287
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发表时间：
2022-09-19
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开发一种化学减毒的、不能复制的病毒疫苗可以提供保护，防止DNA病毒引起的疾病。在这项研究中，我们开发了一种使用centanamycin （CM）生产减毒活病毒的方法，centanamycin （CM）是一种化合物，可以将DNA中富含a - t的小槽烷基化，从而阻止DNA复制。我们测试了CM产生活减毒、复制缺陷的人巨细胞病毒、小鼠巨细胞病毒和单纯疱疹病毒-2 （HSV-2）的功效，这表明CM在产生活减毒、复制缺陷的DNA病毒方面具有广泛的应用。质谱分析表明，CM在腺嘌呤- n3位点烷基化病毒DNA。此外，用cm减毒小鼠巨细胞病毒（MCMV）免疫小鼠可产生强大的免疫应答，并降低免疫动物对活的野生型MCMV的病毒载量。我们的研究提供了一个统一的和有吸引力的治疗机会，化学减毒活DNA病毒可以很容易地开发作为新的一线疫苗。用Centanamycin （CM）处理的DNA病毒免疫可诱导强烈的中和抗体反应，用CM处理的小鼠巨细胞病毒免疫小鼠可免受活病毒的攻击。CM处理的DNA病毒中存在大量独特的病毒抗原，包括主要的人类病原体，如疱疹病毒、腺病毒、乳头瘤病毒、肠病毒等。到目前为止，还没有一种确定的方法可以产生用于疫苗开发的减毒活病毒和复制缺陷病毒。一种可以应用于所有DNA病毒的通用技术为这一问题提供了可行的解决方案。我们提出的技术解决方案是使用centanamycin （CM）来开发全病毒粒子，化学减毒，复制缺陷的活DNA病毒作为潜在的疫苗。我们发现CM在体外能有效抑制人巨细胞病毒（HCMV）、小鼠巨细胞病毒（MCMV）和单纯疱疹病毒-2 （HSV-2）的复制。我们的技术可以在几小时内制造出减毒的DNA复制缺陷病毒。它很简单，可快速扩展，适用于任何类型的DNA病毒，需要较少的劳动力，而且价格低廉。重要的是，我们证明了cm减毒的全病毒粒子MCMV在小鼠中引起了强大的血清中和抗体反应，并且接种疫苗的动物完全免受活病毒的攻击。Jaijyan等人描述了一种生成用于疫苗开发的化学减毒、复制缺陷DNA病毒的有效方法。CM使DNA病毒的基因组烷基化。经免疫后，cm处理的病毒诱导强烈的免疫激活，并提供对活病毒挑战的保护。本方法适用于所有DNA病毒。
The development of a chemically attenuated, replication-incompetent virus vaccine can provide protection against diseases caused by DNA viruses. In this study, we have developed a method to produce live-attenuated, replication-defective viruses using centanamycin (CM), a chemical compound that alkylates the A-T-rich minor groove of the DNA and thereby blocks DNA replication. We tested the efficacy of CM to produce live-attenuated, replication-defective human cytomegalovirus, mouse cytomegalovirus, and herpes simplex virus-2 (HSV-2), suggesting a broad application for generating live-attenuated, replication-defective DNA viruses. Mass spectrometry analysis showed that CM alkylate viral DNA at the adenine-N3 position. Moreover, mice immunization with CM-attenuated mouse cytomegalovirus (MCMV) produced a robust immune response and reduced the viral load in immunized animals against challenges with live, wild-type MCMV. Our study offers a unifying and attractive therapeutic opportunity that chemically attenuated live DNA viruses can be readily developed as new frontline vaccines. Centanamycin (CM) treatment creates live-attenuated, replication-defective DNA viruses Immunization with CM-treated DNA virus induces strong neutralizing antibody response Mice immunized with CM-treated mouse CMV are protected against live virus challenges A large pool of unique viral antigens is present in CM-treated DNA viruses DNA viruses include major human pathogens such as herpesviruses, adenoviruses, papillomaviruses, enteroviruses, and others. As of now, there is not an established method that can generate live-attenuated, replication-defective viruses for vaccine development. A common technology that can be applied to all DNA viruses presents a feasible solution to this problem. Our proposed technology solution is the use of centanamycin (CM) to develop whole-virion, chemically attenuated, replication-defective live DNA viruses as potential vaccines. We found that CM potently inhibits replication of human cytomegalovirus (HCMV), mouse CMV (MCMV), and herpes simplex virus-2 (HSV-2) in vitro. Our technology can produce live-attenuated, DNA replication-defective viruses in a few hours. It is easy, quickly scalable, adaptable to any type of DNA virus, requires less labor, and is inexpensive. Importantly, we demonstrate that CM-attenuated whole virion MCMV elicits a robust serum neutralizing antibody response in mice and that the vaccinated animals were completely protected from live-virus challenge. Jaijyan et al. describe an efficient method to generate chemically live-attenuated, replication-defective DNA viruses for vaccine development. CM alkylates the genome of DNA viruses. Upon immunization, CM-treated viruses induce strong immune activation and provide protection against live virus challenges. The method developed here can be applied to all DNA viruses.
影响因子：
3
作者：
Rayburn E;Wang W;Li M;Zhang X;Xu H;Li H;Qin JJ;Jia L;Covey J;Lee M;Zhang R
通讯作者：
Zhang R