甘露糖修饰的壳聚糖聚乳酸-羟基乙酸共聚物纳米微球作为口蹄疫病毒核酸疫苗递送载体的特性评价

doi:10.11843/j.issn.0366-6964.2021.012.022

Abstract

Abstract: This study aimed to evaluate the characteristics of mannosylated chitosan polylactic acid-glycolide copolymer[poly(D,L-lactide-co-glycolide), PLGA] nanospheres as a delivery system of FMDV nucleic acid vaccine. A certain degree of substitution of mannose modified chitosan (MCS) was prepared by schiff base reaction and elemental analysis. Then, Mannose modified chitosan PLGA nanospheres (MCS-PLGA-NPs) were prepared by double emulsification and volatilization. The particle size distribution and surface potential (zeta) of MCS-PLGA-NPS were measured by nanometer particle size analyzer; The morphology of MCS-PLGA-NPS was investigated by scanning electron microscopy; The ability of plasmids adsorbed on MCS-PLGA-NPs to resist nuclease degradation was observed by agarose gel electrophoresis; The cytotoxicity of MCS-PLGA-NPs was detected by CCK-8 method; The MCS-PLGA-NPs-DNA complex uptake by macrophages was observed by confocal laser scanning; The expression of MCS-PLGA-NPs loaded plasmid DNA in cells was verified by fluorescence microscope and Western blot. The results of elemental analysis showed that MCS with a substitution degree of 5% to 10% was successfully prepared; The results of nanometer particle size measurement and scanning electron microscopy showed that the zeta of MCS-PLGA-NPs was positive, the particle size distribution was uniform and the morphology was spherical; The results of agarose gel electrophoresis showed that the ability of MCS-PLGA-NPs to adsorb plasmids increased with the increase of their quality and could resist nuclease degradation of plasmid DNA to a certain extent; In the cytotoxicity test, the survival rate of RAW264.7 cells was still more than 85% after 24 h co-incubation with different concentrations of MCS-PLGA-NPs; In the cell uptake experiment, confocal laser microscopy could clearly observe the plasmid DNA binding to the surface of the nanomicrosphere and uptake by RAW264.7 cells; Fluorescence microscopy and Western blot experiments showed that MCS-PLGA-NPS-loaded plasmid DNA could be expressed in cells. In summary, MCS and MCS-PLGA-NPs with the ability to deliver DNA vaccine of FMDV were successfully prepared, which provided new directions and insights for the delivery of FMDV nucleic acid vaccine, and also laid a foundation for further research on the carrier carrying specific antigen targeting mannose receptor on APCs surface and its application in animal immunity.

Key words: FMDV, mannose modified chitosan, chitosan grafted PLGA, nanospheres, delivery system, DNA vaccine

CLC Number:

Q933
S855.3

LI Xian, ZHANG Zhongwang, ZHANG Fudong, Lü Jianliang, LI Jiahao, PAN Li. Evaluation of the Characteristics of Mannosylated Chitosan PLGA Nanospheres as a Delivery System for DNA Vaccine of A/FMDV[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3557-3568.

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Evaluation of the Characteristics of Mannosylated Chitosan PLGA Nanospheres as a Delivery System for DNA Vaccine of A/FMDV

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