Laboratorio de Biofarmacéuticos Recombinantes, Facultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava 6, 78210, San Luis Potosí, SLP, Mexico,
Plant Cell Rep. 2015 Mar;34(3):425-33. doi: 10.1007/s00299-014-1720-6. Epub 2014 Dec 5.
The first report on the recombinant production of a candidate vaccine in the moss system. The need for economical and efficient platforms for vaccine production demands the exploration of emerging host organisms. In this study, the production of an antigenic protein is reported employing the moss Physcomitrella patens as an expression host. A multi-epitope protein from the Human Immunodeficiency Virus (HIV) based on epitopes from gp120 and gp41 was designed as a candidate subunit vaccine and named poly-HIV. Transgenic moss plants were generated carrying the corresponding poly-HIV transgene under a novel moss promoter and subsequently seven positive lines were confirmed by PCR. The poly-HIV protein accumulated up to 3.7 µg g(-1) fresh weight in protonema cultures. Antigenic and immunogenic properties of the moss-produced recombinant poly-HIV are evidenced by Western blots and by mice immunization assays. The elicitation of specific antibodies in mice was observed, reflecting the immunogenic potential of this moss-derived HIV antigen. This is the first report on the production of a potential vaccine in the moss system and opens the avenue for glycoengineering approaches for the production of HIV human-like glycosylated antigens as well as other vaccine prototypes under GMP conditions in moss bioreactors.
首个关于在苔藓系统中重组生产候选疫苗的报告。为了满足经济高效的疫苗生产需求,需要探索新兴的宿主生物。在这项研究中,我们利用苔藓植物Physcomitrella patens 作为表达宿主,生产一种抗原蛋白。一种基于 HIV 包膜糖蛋白 gp120 和 gp41 表位的多表位蛋白被设计为候选亚单位疫苗,并命名为 poly-HIV。携带相应 poly-HIV 转基因的转基因苔藓植物在一个新型苔藓启动子的控制下生成,并通过 PCR 证实了七个阳性株系。在原丝体培养物中,poly-HIV 蛋白积累量高达 3.7μg/g 鲜重。通过 Western blot 和小鼠免疫实验证实了苔藓产生的重组 poly-HIV 的抗原性和免疫原性。在小鼠中观察到了特异性抗体的产生,反映了这种苔藓衍生的 HIV 抗原的免疫原性潜力。这是首个关于在苔藓系统中生产潜在疫苗的报告,为在苔藓生物反应器中生产具有人类样糖基化抗原的 HIV 以及其他疫苗原型的糖基工程方法开辟了道路。