Key Laboratory of Bio-resources and Eco-environment (Ministry of Education), College of Life Sciences, Sichuan University, Chengdu, China.
Sichuan Engineering Technology Research Center of Medical Animal, Chengdu, China.
J Nanobiotechnology. 2018 Jan 29;16(1):8. doi: 10.1186/s12951-018-0337-2.
Trueperella pyogenes is a commensal and opportunistic pathogen that normally causes mastitis, liver abscesses and pneumonia of economically important livestock. To develop efficacious and potent vaccine against T. pyogenes, chimeric gene DNA vaccines were constructed and encapsulated in chitosan nanoparticles (pPCFN-CpG-CS-NPs).
The pPCFN-CpG-CS-NPs consists of the plo, cbpA, fimA, and nanH gene of T. pyogenes and CpG ODN1826. It was produced with good morphology, high stability, a mean diameter of 93.58 nm, and a zeta potential of + 5.27 mV. Additionally, chitosan encapsulation was confirmed to protect the DNA plasmid from DNase I digestion. The immunofluorescence assay indicated that the four-chimeric gene could synchronously express in HEK293T cells and maintain good bioactivity. Compared to the mice immunized with the control plasmid, in vivo immunization showed that mice immunized with the pPCFN-CpG-CS-NPs had better immune responses, and release of the plasmid DNA was prolonged. Importantly, immunization with pPCFN-CpG-CS-NPs could significantly protect mice from highly virulent T. pyogenes TP7 infection.
This study indicates that chitosan-DNA nanoparticles are potent immunization candidates against T. pyogenes infection and provides strategies for the further development of novel vaccines encapsulated in chitosan nanoparticles.
化脓隐秘杆菌是一种共生和机会性病原体,通常会导致经济重要家畜的乳腺炎、肝脓肿和肺炎。为了开发针对化脓隐秘杆菌的有效和强效疫苗,构建了嵌合基因 DNA 疫苗并封装在壳聚糖纳米粒子(pPCFN-CpG-CS-NPs)中。
pPCFN-CpG-CS-NPs 由化脓隐秘杆菌的 plo、cbpA、fimA 和 nanH 基因和 CpG ODN1826 组成。它具有良好的形态、高稳定性、平均直径为 93.58nm 和 +5.27mV 的 ζ 电位。此外,壳聚糖包封被证实可以保护 DNA 质粒免受 DNase I 消化。免疫荧光分析表明,四个嵌合基因可以在 HEK293T 细胞中同步表达并保持良好的生物活性。与用对照质粒免疫的小鼠相比,体内免疫显示用 pPCFN-CpG-CS-NPs 免疫的小鼠具有更好的免疫反应,并且质粒 DNA 的释放得到延长。重要的是,pPCFN-CpG-CS-NPs 的免疫接种可以显著保护小鼠免受高毒力 T. pyogenes TP7 感染。
本研究表明,壳聚糖-DNA 纳米粒子是针对化脓隐秘杆菌感染的有效免疫候选物,并为进一步开发壳聚糖纳米粒子包封的新型疫苗提供了策略。