Rodrigues-Jesus M J, Fotoran W L, Cardoso R M, Araki K, Wunderlich G, Ferreira Luís C S
Vaccine Development Laboratory, Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, Av. Prof. Lineu Prestes, 1374, Cidade Universitária, São Paulo, SP, 05508-900, Brazil.
Unit for Drug Development and Plasmodium Molecular Biology, Department of Parasitology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
Braz J Microbiol. 2019 Jan;50(1):67-77. doi: 10.1007/s42770-018-0035-0. Epub 2018 Dec 6.
Microlipid vesicles (MLV) have a broad spectrum of applications for the delivery of molecules, ranging from chemical compounds to proteins, in both in vitro and in vivo conditions. In the present study, we developed a new set of nanosize multilayer lipid vesicles (NMVs) containing a unique combination of lipids. The NMVs enable the adsorption of histidine-tagged proteins at the vesicle surface and were demonstrated to be suitable for the in vivo delivery of antigens. The NMVs contained a combination of neutral (DOPC) and anionic (DPPG) lipids in the inner membrane and an external layer composed of DOPC, cholesterol, and a nickel-containing lipid (DGS-NTA [Ni]). NMVs combined with a recombinant form of the B subunit of the Shiga toxin (rStx2B) produced by certain enterohemorragic Escherichia coli (EHEC) strains enhanced the immunogenicity of the antigen after parenteral administration to mice. Mice immunized with rStx2B-loaded NMVs elicited serum antibodies capable of neutralizing the toxic activities of the native toxin; this result was demonstrated both in vitro and in vivo. Taken together, these results demonstrated that the proposed NMVs represent an alternative for the delivery of antigens, including recombinant proteins, generated in different expression systems.
微脂质体（MLV）在体外和体内条件下，对于从化合物到蛋白质等各种分子的递送都有广泛的应用。在本研究中，我们开发了一组新的纳米级多层脂质体（NMV），其含有独特的脂质组合。NMV能够使组氨酸标签蛋白吸附在囊泡表面，并被证明适用于抗原的体内递送。NMV的内膜含有中性脂质（二油酰磷脂酰胆碱，DOPC）和阴离子脂质（二棕榈酰磷脂酰甘油，DPPG）的组合，外层由DOPC、胆固醇和含镍脂质（二硬脂酰基磷脂酰乙醇胺 - N - 三（羧甲基）氨基甲酰基 - N - （镍），DGS - NTA [Ni]）组成。与某些肠出血性大肠杆菌（EHEC）菌株产生的志贺毒素B亚基重组形式（rStx2B）结合的NMV，在经肠胃外给药小鼠后增强了抗原的免疫原性。用负载rStx2B的NMV免疫的小鼠产生了能够中和天然毒素毒性活性的血清抗体；这一结果在体外和体内均得到证实。综上所述，这些结果表明，所提出的NMV代表了一种用于递送包括在不同表达系统中产生的重组蛋白在内的抗原的替代方法。
