Pasandideh Reza, Seyfi Abad Shapouri Masoud Reza, Beigi Nassiri Mohammad Taghi
Department of Animal Science, Khuzestan Agricultural Sciences and Natural Resources University, Ahvaz.
Onderstepoort J Vet Res. 2018 Aug 28;85(1):e1-e6. doi: 10.4102/ojvr.v85i1.1617.
The aim of this study was to investigate the immunogenicity of a plasmid deoxyribonucleic acid (DNA) vaccine encoding the G1 epitope of bovine ephemeral fever virus (BEFV) G glycoprotein in mice. A plasmid DNA carrying the G1 gene was constructed and designated as pcDNA3.1-G1. The expression of the target gene was confirmed in human embryonic kidney 293 (HEK 293) cells transfected with pcDNA3.1-G1 by indirect immunofluorescent staining. Immunisation experiments were intramuscularly carried out by vaccinating 6-week-old female mice in four groups, including the pcDNA3.1-G1 construct, pcDNA3.1 (+) plasmid alone, BEF-inactivated vaccine and phosphate-buffered saline (PBS) (1X) three times with 2-week intervals. Fourteen days after the last immunisation, the animals were bled and the resulting sera were tested for anti-G1-specific antibodies by immunoblotting analysis, indirect enzyme-linked immunosorbent assay (ELISA) and virus neutralisation (VN) test. Serological assays showed that the pcDNA3.1-G1 construct expressing G1 protein was able to elicit specific antibodies against this antigen. Virus neutralisation test showed that pcDNA3.1-G1 could induce anti-BEFV-neutralising antibodies in mice. Our findings indicated that a new dimension can be added to vaccine studies for bovine ephemeral fever (BEF) using eukaryotic expression plasmids encoding the G1 antigen in the future.
本研究旨在探讨编码牛暂时热病毒(BEFV)G糖蛋白G1表位的质粒脱氧核糖核酸(DNA)疫苗在小鼠体内的免疫原性。构建了携带G1基因的质粒DNA,并命名为pcDNA3.1-G1。通过间接免疫荧光染色在转染了pcDNA3.1-G1的人胚肾293(HEK 293)细胞中证实了靶基因的表达。免疫实验通过对6周龄雌性小鼠进行肌肉注射来进行,分为四组,包括pcDNA3.1-G1构建体、单独的pcDNA3.1(+)质粒、BEF灭活疫苗和磷酸盐缓冲盐水(PBS)(1X),每隔2周接种3次。最后一次免疫后14天,采集动物血液,通过免疫印迹分析、间接酶联免疫吸附测定(ELISA)和病毒中和(VN)试验检测所得血清中的抗G1特异性抗体。血清学检测表明,表达G1蛋白的pcDNA3.1-G1构建体能够引发针对该抗原的特异性抗体。病毒中和试验表明,pcDNA3.1-G1可在小鼠体内诱导抗BEFV中和抗体。我们的研究结果表明,未来使用编码G1抗原的真核表达质粒对牛暂时热(BEF)疫苗研究可增添新的维度。