Institute Science Biology, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia.
Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
Mol Biotechnol. 2020 May;62(5):289-296. doi: 10.1007/s12033-020-00244-0.
Pasteurella multocida is the main cause of haemorrhagic septicaemia (HS) outbreak in livestock, such as cattle and buffaloes. Conventional vaccines such as alum-precipitated or oil-adjuvant broth bacterins were injected subcutaneously to provide protection against HS. However, the immunity developed is only for short term and needed to be administered frequently. In our previous study, a short gene fragment from Pasteurella multocida serotype B was obtained via shotgun cloning technique and later was cloned into bacterial expression system. pQE32-ABA392 was found to possess immunogenic activity towards HS when tested in vivo in rat model. In this study, the targeted gene fragment of ABA392 was sub-cloned into a DNA expression vector pVAX1 and named as pVAX1-ABA392. The new recombinant vaccine was stable and expressed on mammalian cell lines. Serum sample collected from a group of vaccinated rats for ELISA test shows that the antibody in immunized rats was present at high titer and can be tested as a vaccine candidate with challenge in further studies. This successful recombinant vaccine is immunogenic and potentially could be used as vaccine in future against HS.
多杀巴斯德氏菌是牛、水牛等牲畜发生出血性败血症(HS)的主要原因。常规疫苗,如明矾沉淀或油佐剂菌苗,通过皮下注射提供针对 HS 的保护。然而,产生的免疫力只是短期的,需要频繁给药。在我们之前的研究中,通过鸟枪法克隆技术获得了多杀巴斯德氏菌 B 型的一个短基因片段,后来将其克隆到细菌表达系统中。pQE32-ABA392 在大鼠模型中进行体内试验时被发现具有针对 HS 的免疫原性。在本研究中,ABA392 的目的基因片段被亚克隆到 DNA 表达载体 pVAX1 中,并命名为 pVAX1-ABA392。新型重组疫苗在哺乳动物细胞系中稳定表达。从一组接种疫苗的大鼠中采集的血清样本进行 ELISA 检测显示,免疫大鼠中的抗体呈高滴度存在,并且可以作为候选疫苗进行进一步的挑战试验。这种成功的重组疫苗具有免疫原性,将来可能被用作针对 HS 的疫苗。