Plant-derived protein bodies as delivery vehicles for recombinant proteins into mammalian cells
Plant-derived protein bodies as delivery vehicles for recombinant proteins into mammalian cells
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DOI：
10.1002/bit.27273
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发表时间：
2020-01-30
影响因子：
3.8
通讯作者：
Stoger, Eva
中科院分区：
文献类型：
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作者：
Schwestka, Jennifer;Tschofen, Marc;Stoger, Eva
将生物药物包裹成微米或纳米颗粒是一种经常用于防止降解或实现治疗药物和疫苗缓慢释放的策略。蛋白质小体(PBS)是种子中天然存在的储存细胞器，可以作为载体。玉米贮藏蛋白的N-末端序列与其他蛋白质的融合足以诱导PBS的形成，PBS可用于将重组蛋白直接生物包裹在植物生产宿主中。此外，玉米醇溶蛋白的免疫刺激作用也已有报道，这有利于疫苗的投放。然而，对玉米醇溶蛋白PBS与哺乳动物细胞之间的相互作用知之甚少。为了更好地了解这种相互作用，通过将玉米醇溶蛋白的N端部分与绿色荧光蛋白融合而产生的荧光PBS，在本氏烟草叶片中产生，通过基于过滤的下游程序回收，并用于研究它们在哺乳动物细胞中的内化效率。我们发现，与同等大小的聚苯乙烯微珠相比，荧光PBS有效地内化到肠上皮细胞和抗原提呈细胞(APC)中。此外，我们观察到PBS刺激上皮细胞分泌细胞因子，这一特征可能通过招募APC来赋予疫苗佐剂活性。综上所述，这些结果支持使用玉米醇溶蛋白融合蛋白来开发基于受控蛋白包装到植物PBS中的药物输送的新方法。
The encapsulation of biopharmaceuticals into micro- or nanoparticles is a strategy frequently used to prevent degradation or to achieve the slow release of therapeutics and vaccines. Protein bodies (PBs), which occur naturally as storage organelles in seeds, can be used as such carrier vehicles. The fusion of the N-terminal sequence of the maize storage protein, gamma-zein, to other proteins is sufficient to induce the formation of PBs, which can be used to bioencapsulate recombinant proteins directly in the plant production host. In addition, the immunostimulatory effects of zein have been reported, which are advantageous for vaccine delivery. However, little is known about the interaction between zein PBs and mammalian cells. To better understand this interaction, fluorescent PBs, resulting from the fusion of the N-terminal portion of zein to a green fluorescent protein, was produced in Nicotiana benthamiana leaves, recovered by a filtration-based downstream procedure, and used to investigate their internalization efficiency into mammalian cells. We show that fluorescent PBs were efficiently internalized into intestinal epithelial cells and antigen-presenting cells (APCs) at a higher rate than polystyrene beads of comparable size. Furthermore, we observed that PBs stimulated cytokine secretion by epithelial cells, a characteristic that may confer vaccine adjuvant activities through the recruitment of APCs. Taken together, these results support the use of zein fusion proteins in developing novel approaches for drug delivery based on controlled protein packaging into plant PBs.