Kamal Nuriqmaliza M, Zamri-Saad M, Masarudin Mas Jaffri, Othman Sarah
Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia.
Research Centre for Ruminant Diseases, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia.
BMC Vet Res. 2017 Jun 19;13(1):186. doi: 10.1186/s12917-017-1109-1.
Pasteurella multocida B:2 causes bovine haemorrhagic septicaemia (HS), leading to rapid fatalities in cattle and buffaloes. An attenuated derivative of P. multocida B:2 GDH7, was previously constructed through mutation of the gdhA gene and proved to be an effective live attenuated vaccine for HS. Currently, only two potential live attenuated vaccine candidates for HS are being reported; P. multocida B:2 GDH7 and P. multocida B:2 JRMT12. This study primarily aims to investigate the potential of P. multocida B:2 GDH7 strain as a delivery vehicle for DNA vaccine for future multivalent applications.
An investigation on the adherence, invasion and intracellular survival of bacterial strains within the bovine aortic endothelial cell line (BAEC) were carried out. The potential vaccine strain, P. multocida B:2 GDH7, was significantly better (p ≤ 0.05) at adhering to and invading BAEC compared to its parent strain and to P. multocida B:2 JRMT12 and survived intracellularly 7 h post treatment, with a steady decline over time. A dual reporter plasmid, pSRGM, which enabled tracking of bacterial movement from the extracellular environment into the intracellular compartment of the mammalian cells, was subsequently transformed into P. multocida B:2 GDH7. Intracellular trafficking of the vaccine strain, P. multocida B:2 GDH7 was subsequently visualized by tracking the reporter proteins via confocal laser scanning microscopy (CLSM).
The ability of P. multocida B:2 GDH7 to model bactofection represents a possibility for this vaccine strain to be used as a delivery vehicle for DNA vaccine for future multivalent protection in cattle and buffaloes.
多杀性巴氏杆菌B:2型可引发牛出血性败血症（HS），导致牛和水牛迅速死亡。多杀性巴氏杆菌B:2型GDH7的减毒株先前通过gdhA基因突变构建而成，被证明是一种有效的HS活疫苗。目前，仅有两种潜在的HS活疫苗候选株被报道，即多杀性巴氏杆菌B:2型GDH7和多杀性巴氏杆菌B:2型JRMT12。本研究主要旨在探究多杀性巴氏杆菌B:2型GDH7菌株作为DNA疫苗递送载体用于未来多价应用的潜力。
对牛主动脉内皮细胞系（BAEC）内细菌菌株的黏附、侵袭和细胞内存活情况进行了研究。与亲本菌株以及多杀性巴氏杆菌B:2型JRMT12相比，潜在疫苗株多杀性巴氏杆菌B:2型GDH7在黏附并侵袭BAEC方面表现显著更优（p≤0.05），且在处理后7小时内存活于细胞内，随时间稳定下降。随后将一种能够追踪细菌从细胞外环境进入哺乳动物细胞内区室移动的双报告质粒pSRGM转化到多杀性巴氏杆菌B:2型GDH7中。随后通过共聚焦激光扫描显微镜（CLSM）追踪报告蛋白，观察到疫苗株多杀性巴氏杆菌B:2型GDH7在细胞内的转运情况。
多杀性巴氏杆菌B:2型GDH7模拟细菌转染的能力表明，该疫苗株有可能用作DNA疫苗的递送载体，用于未来牛和水牛的多价保护。
