Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, Rondebosch, 7701, South Africa.
Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, Rondebosch, 7701, South Africa; Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Rondebosch, 7701, South Africa.
Virology. 2023 Mar;580:88-97. doi: 10.1016/j.virol.2023.02.003. Epub 2023 Feb 12.
Human papillomaviruses (HPVs) are known to be the cause of anogenital and oropharyngeal cancers as well as genital and common warts. HPV pseudovirions (PsVs) are synthetic viral particles that are made up of the L1 major and L2 minor HPV capsid proteins and up to 8 Kb of encapsidated pseudogenome dsDNA. HPV PsVs are used to test novel neutralising antibodies elicited by vaccines, for studying the virus life cycle, and potentially for the delivery of therapeutic DNA vaccines. HPV PsVs are typically produced in mammalian cells, however, it has recently been shown that Papillomavirus PsVs can be produced in plants, a potentially safer, cheaper and more easily scalable means of production. We analysed the encapsidation frequencies of pseudogenomes expressing EGFP, ranging in size from 4.8 Kb to 7.8 Kb, by plant-made HPV-35 L1/L2 particles. The smaller pseudogenomes were found to be packaged more efficiently into PsVs as higher concentrations of encapsidated DNA and higher levels of EGFP expression were obtained with the 4.8 Kb pseudogenome, compared to the larger 5.8-7.8 Kb pseudogenomes. Thus, smaller pseudogenomes, of 4.8 Kb, should be used for efficient plant production of HPV-35 PsVs.
人乳头瘤病毒（HPV）是已知的肛门生殖器和口咽癌以及生殖器和常见疣的病因。HPV 假病毒（PsV）是由 L1 主要和 L2 次要 HPV 衣壳蛋白以及最多 8kb 包裹的假基因组 dsDNA 组成的合成病毒颗粒。HPV PsV 用于测试疫苗引起的新型中和抗体，用于研究病毒生命周期，并且具有潜在的用于传递治疗性 DNA 疫苗的用途。HPV PsV 通常在哺乳动物细胞中产生，然而，最近已经表明，Papillomavirus PsV 可以在植物中产生，这是一种更安全、更便宜且更容易扩展的生产方式。我们通过植物制造的 HPV-35 L1/L2 颗粒分析了大小在 4.8kb 至 7.8kb 之间的表达 EGFP 的假基因组的包装频率。较小的假基因组被发现更有效地包装到 PsV 中，因为与较大的 5.8-7.8kb 假基因组相比，获得了更高浓度的包裹 DNA 和更高水平的 EGFP 表达。因此，较小的假基因组（4.8kb）应用于高效的植物生产 HPV-35 PsV。
