Preparation and efficacy of Newcastle disease virus DNA vaccine encapsulated in chitosan nanoparticles

Abstract

Optimal preparation conditions of Newcastle disease virus (NDV) F gene deoxyribonucleic acid (DNA) vaccine encapsulated in chitosan nanoparticles (pFNDV-CS-NPs) were determined. The pFNDV-CS-NPs were prepared according to a complex coacervation method. The pFNDV-CS-NPs were produced with good morphology, high stability, a mean diameter of 199.5 nm, encapsulation efficiency of 98.37%±0.87%, loading capacity of 36.12%±0.19%, and a zeta potential of +12.11 mV. The in vitro release assay showed that the plasmid DNA was sustainably released from the pFNDV-CS-NPs, up to 82.9%±2.9% of the total amount. Cell transfection test indicated that the vaccine expressed the F gene in cells and maintained good bioactivity. Additionally, the safety of mucosal immunity delivery system of the pFNDV-CS-NPs was also tested in vitro by cell cytotoxicity and in vivo by safety test in chickens. In vivo immunization showed that better immune responses of specific pathogen-free chickens immunized with the pFNDV-CS-NPs were induced, and prolonged release of the plasmid DNA was achieved compared to the chickens immunized with the control plasmid. This study lays the foundation for the further development of mucosal vaccines and drugs encapsulated in chitosan nanoparticles.

Synthesis, characterization, and immune efficacy of layered double hydroxide@SiO2 nanoparticles with shell-core structure as a delivery carrier for Newcastle disease virus DNA vaccine

“…36 The particle size and zeta potentials of both pFDNA-SiO 2 -NPs and pFDNA-LDH@SiO 2 -NPs were measured using a Zeta Sizer 2000 from Malvern Instruments (Malvern, UK). The loading capacity (LC) of the pFDNA-LDH@SiO 2 -NPs was calculated as follows:…”

“…36,38 Chicken embryo kidney cells were first cultured in DMEM, then diluted to the final cell concentration of 2×10 6 cells/mL, transferred to 96-well plates at 200 µL per well, and cultured at 37°C for 5 hours. After discarding the supernatant in the wells, 50 µL of pFDNA-LDH@SiO 2 -NPs (diluted in DMEM culture at 1 mg/mL) was added into the wells, followed by incubation at 37°C for 2 hours.…”

“…36 safety of the pFDNa-lDh@siO 2 -NPs Fifteen 7-day-old SPF chickens obtained from Harbin Veterinary Research Institute Laboratory Animal Center (Harbin, People's Republic of China) were randomly divided into three groups: chickens in group 1 were immunized intramuscularly (IM) with the pFDNA-LDH@SiO 2 -NPs containing 2 mg of the plasmid DNA; chickens in group 2 were immunized intranasally (IN) with the pFDNA-LDH@SiO 2 -NPs containing 2 mg of the plasmid DNA; and chickens in group 3 were immunized IM with the PBS solution as control group. These three groups of chickens were bred under the same conditions.…”

“…Protection was determined as described previously. 36 Five chickens were randomly selected from every immunized group when the level of NDV-specific antibody in serum was increased to 6.0 log 2. The chickens infected IM with 0.1 mL of the highly virulent NDV strain F 48 E 9 had a viral titer of 10 4.5 50% egg infectious dose/0.1 mL for challenge studies.…”

Newcastle disease virus vaccine encapsulated in biodegradable nanoparticles for mucosal delivery of a human vaccine

“…Wei Li et al 13 used the method of double emulsion-solvent evaporation to prepare PLGA-plasmid DNA nanoparticles (pFNDV-PLGANPs) with the advantage of sustained release effect and perfect immune effect. Zhang et al 21 prepared F gene plasmid DNA of NDV encapsulated in chitosan nanoparticles (pFNDV-CS-NPs) based on complex coacervation. Gang Chen et al 14 also did interrelated work and prepared novel mucosal delivery system as chitosan nanoparticles containing the lentogenic live-virus vaccine (NDV-CS-NPs) using ionic cross linking method.…”