Chockalingam A K, Thiyagarajan S, Govindasamy N, Patnaikuni R, Garlapati S, Golla R R, Joyappa D H, Krishnamshetty P, Veluvarti V V S
Indian Veterinary Research Institute, Bangalore, India.
Acta Virol. 2010;54(3):189-95. doi: 10.4149/av_2010_03_189.
Since foot-and-mouth disease virus (FMDV) serotypes display a great genetic and antigenic diversity, there is a constant requirement to monitor the performance of FMDV vaccines in the field with respect to their antigenic coverage. To avoid possible antigenic changes in field FMDV isolates during their adaptation to BHK-21 cells, a standard step used in production of conventional FMDV vaccines, the custom-made chimeric conventional or DNA vaccines, in which antigenic determinants are replaced with those of appropriate field strains, should be constructed. Using this approach, we made a plasmid-based chimeric FMDV DNA vaccine containing structural genes of serotype O in the genome backbone of serotype Asia 1, all under the control of Human cytomegalovirus (HCMV) immediate early gene promoter. BHK-21 cells transfected with the chimeric DNA vaccine did not show cytopathic effect (CPE), but expressed virus-specific proteins as demonstrated by 35S-methionine labeling and immunoprecipitation. Guinea pigs immunized with the chimeric DNA vaccine produced virus-specific antibodies assayed by ELISA and virus neutralization test (VNT), respectively. The chimeric DNA vaccine showed a partial protection of guinea pigs challenged with the virulent FMDV. Although the chimeric DNA vaccine, in general, was not as effective as a conventional one, this study encourages further work towards the development of genetically engineered custom-made chimeric vaccines against FMDV.
由于口蹄疫病毒(FMDV)血清型具有很大的遗传和抗原多样性,因此持续需要监测FMDV疫苗在田间的抗原覆盖性能。为避免在传统FMDV疫苗生产中使用的标准步骤,即田间FMDV分离株在适应BHK - 21细胞过程中可能出现的抗原变化,应构建定制的嵌合传统疫苗或DNA疫苗,其中抗原决定簇被适当的田间毒株的抗原决定簇所取代。采用这种方法,我们构建了一种基于质粒的嵌合FMDV DNA疫苗,其在亚洲1型血清型的基因组骨架中包含O型血清型的结构基因,所有这些都在人巨细胞病毒(HCMV)立即早期基因启动子的控制下。用嵌合DNA疫苗转染的BHK - 21细胞未显示细胞病变效应(CPE),但如35S - 甲硫氨酸标记和免疫沉淀所示,表达了病毒特异性蛋白。分别用ELISA和病毒中和试验(VNT)检测,用嵌合DNA疫苗免疫的豚鼠产生了病毒特异性抗体。嵌合DNA疫苗对用强毒FMDV攻击的豚鼠显示出部分保护作用。虽然嵌合DNA疫苗总体上不如传统疫苗有效,但这项研究鼓励进一步开展针对FMDV的基因工程定制嵌合疫苗的研发工作。