ICAR-Indian Veterinary Research Institute, Hebbal, Bengaluru 560024, India.
ICAR-Indian Veterinary Research Institute, Hebbal, Bengaluru 560024, India.
Int J Biol Macromol. 2024 Oct;277(Pt 4):134366. doi: 10.1016/j.ijbiomac.2024.134366. Epub 2024 Aug 3.
Intact capsids of foot-and-mouth disease virus (FMDV) play a vital role in eliciting a protective immune response. Any change in the physico-chemical environment of the capsids results in dissociation and poor immunogenicity. Structural bioinfomatics studies have been carried out to predict the amino acids at the interpentameric region that resulted in the identification of mutant virus-like particles(VLPs) of FMDV serotype Asia1/IND/63/1972. The insect cell expressed VLPs were evaluated for their stability by sandwich ELISA. Among 10 mutants, S93H showed maximum retention of antigenicity at different temperatures, indicating its higher thermal stability as revealed by the in-silico analysis and retained the antigenic sites of the virus demonstrated by Sandwich ELISA. The concordant results of the liquid phase blocking ELISA for estimation of antibody titre of known sera with stable mutant VLP as antigen in place of virus antigen demonstrate its diagnostic potential. The stable mutant VLP elicited a robust immune response with 85.6 % protection in guinea pigs against virus challenge. The stabilized VLP based antigen requires minimum biosafety and cold storage for production and transit besides, complying with differentiation of infected from vaccinated animals. It can effectively replace the conventional virus handling during antigen production for prophylactic and diagnostic use.
口蹄疫病毒(FMDV)完整的衣壳在引发保护性免疫反应方面起着至关重要的作用。衣壳的理化环境的任何变化都会导致衣壳的解离和免疫原性降低。结构生物信息学研究已经被用来预测在五聚体间区域导致鉴定出 FMDV 血清型 Asia1/IND/63/1972 的突变病毒样颗粒(VLPs)的氨基酸。通过夹心 ELISA 评估了在昆虫细胞中表达的 VLPs 的稳定性。在 10 个突变体中,S93H 在不同温度下表现出最大的抗原保留性,表明其具有更高的热稳定性,这是通过计算机分析和保留病毒抗原的夹心 ELISA 揭示的抗原表位来证实的。在液相阻断 ELISA 中,用稳定的突变 VLP 代替病毒抗原作为抗原,对已知血清的抗体滴度进行估计的结果是一致的,这表明它具有诊断潜力。稳定的突变 VLP 能够在豚鼠中引发强烈的免疫反应,对病毒攻击的保护率达到 85.6%。基于稳定的 VLP 的抗原在生产和运输过程中需要最低的生物安全和冷藏要求,并且符合区分感染动物和接种疫苗动物。它可以有效地替代传统的病毒处理,用于预防和诊断用途。