Pomatto Margherita A C, Gai Chiara, Negro Federica, Massari Lucia, Deregibus Maria Chiara, Grange Cristina, De Rosa Francesco Giuseppe, Camussi Giovanni
EvoBiotech s.r.l., 10122 Turin, Italy.
Department of Medical Science, University of Turin, A.O.U. Città della Salute e della Scienza di Torino, 10126 Turin, Italy.
Pharmaceutics. 2023 Mar 17;15(3):974. doi: 10.3390/pharmaceutics15030974.
Plant-derived extracellular vesicles (EVs) may represent a platform for the delivery of RNA-based vaccines, exploiting their natural membrane envelope to protect and deliver nucleic acids. Here, EVs extracted from orange () juice (oEVs) were investigated as carriers for oral and intranasal SARS-CoV-2 mRNA vaccine. oEVs were efficiently loaded with different mRNA molecules (coding N, subunit 1 and full S proteins) and the mRNA was protected from degrading stress (including RNase and simulated gastric fluid), delivered to target cells and translated into protein. APC cells stimulated with oEVs loaded with mRNAs induced T lymphocyte activation in vitro. The immunization of mice with oEVs loaded with S1 mRNA via different routes of administration including intramuscular, oral and intranasal stimulated a humoral immune response with production of specific IgM and IgG blocking antibodies and a T cell immune response, as suggested by IFN-γ production by spleen lymphocytes stimulated with S peptide. Oral and intranasal administration also triggered the production of specific IgA, the mucosal barrier in the adaptive immune response. In conclusion, plant-derived EVs represent a useful platform for mRNA-based vaccines administered not only parentally but also orally and intranasally.
植物来源的细胞外囊泡（EVs）可能代表了一个用于递送基于RNA的疫苗的平台，利用其天然的膜包膜来保护和递送核酸。在这里，研究了从橙汁中提取的EVs（oEVs）作为口服和鼻内给药的严重急性呼吸综合征冠状病毒2（SARS-CoV-2）mRNA疫苗的载体。oEVs能有效地装载不同的mRNA分子（编码N蛋白、亚基1和完整的S蛋白），并且mRNA能免受降解应激（包括核糖核酸酶和模拟胃液）的影响，被递送至靶细胞并翻译成蛋白质。用装载有mRNA的oEVs刺激的抗原呈递细胞（APC）在体外诱导T淋巴细胞活化。通过包括肌肉内、口服和鼻内在内的不同给药途径，用装载有S1 mRNA的oEVs免疫小鼠，刺激了体液免疫反应，产生了特异性IgM和IgG阻断抗体以及T细胞免疫反应，脾脏淋巴细胞用S肽刺激产生的γ干扰素（IFN-γ）表明了这一点。口服和鼻内给药还触发了特异性IgA的产生，IgA是适应性免疫反应中的黏膜屏障。总之，植物来源的EVs代表了一个不仅可用于肠胃外给药，还可用于口服和鼻内给药的基于mRNA的疫苗的有用平台。
