Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia.
Bioinformatics Programme, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia.
Trop Biomed. 2021 Mar 1;38(1):36-41. doi: 10.47665/tb.38.1.007.
Chikungunya virus (CHIKV) infection is the cause of acute symptoms and chronic symmetrical polyarthritis associated with long-term morbidity and mortality. Currently, there is no available licensed vaccine or particularly useful drug for human use against CHIKV infection. This study was conducted to evaluate the efficacy of antibodies produced by papaya mosaic virus (PapMV) nanoparticles fused to E2EP3 peptide of CHIKV envelope as a recombinant CHIKV vaccine. PapMV, PapMV-C- E2EP3, and E2EP3-N-PapMV were produced in E. coli with an approximate size of 27 to 30 kDa. ICR mice (5 to 6 weeks of age) were injected subcutaneously with 25 micrograms of vaccine construct, and ELISA measured the titer of CHIKV specific IgG antibodies. The results showed that both recombinant proteins E2EP3-N-PapMV and PapMVC-E2EP3 were able to induce IgG antibodies production in immunized mice against CHIKV while immunization with recombinant PapMV showed no IgG antibodies induction. The neutralizing activity of the antibodies generated by either E2EP3-N-PapMV or PapMV-C-E2EP3 exhibited similar inhibition to CHIKV replication in Vero cells using the cells based antibody neutralizing assay and analyzed by plaque formation assay. This study showed the effectiveness of nanoparticles vaccine generated by fusing epitope peptide of CHIKV envelope to papaya mosaic virus envelope in inducing a robust immune response in mice against CHIKV. The data showed that levels of neutralizing antibodies correlate with a protective immune response CHIKV replication.
基孔肯雅病毒（CHIKV）感染是引起急性症状和慢性对称性多关节炎的原因，与长期发病率和死亡率有关。目前，尚无针对 CHIKV 感染的可用许可疫苗或特别有用的人类药物。本研究旨在评估木瓜花叶病毒（PapMV）纳米颗粒与 CHIKV 包膜的 E2EP3 肽融合产生的抗体作为重组 CHIKV 疫苗的功效。PapMV、PapMV-C-E2EP3 和 E2EP3-N-PapMV 在大肠杆菌中产生，大小约为 27 至 30 kDa。ICR 小鼠（5 至 6 周龄）经皮下注射 25 微克疫苗构建体，ELISA 测量针对 CHIKV 的特异性 IgG 抗体滴度。结果表明，两种重组蛋白 E2EP3-N-PapMV 和 PapMVC-E2EP3 均能诱导免疫小鼠产生针对 CHIKV 的 IgG 抗体，而免疫重组 PapMV 则不能诱导 IgG 抗体产生。用基于细胞的抗体中和测定法和蚀斑形成测定法分析，用 E2EP3-N-PapMV 或 PapMV-C-E2EP3 产生的抗体的中和活性均显示出对 Vero 细胞中 CHIKV 复制的相似抑制作用。这项研究表明，将 CHIKV 包膜的表位肽融合到木瓜花叶病毒包膜上产生的纳米颗粒疫苗在诱导小鼠针对 CHIKV 的强大免疫反应方面是有效的。数据表明，中和抗体的水平与针对 CHIKV 复制的保护性免疫反应相关。
