Key Laboratory for Tropic Diseases Control, Ministry of Education of China, Department of Microbiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, Guangdong, China.
Department of Gynaecology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou City, Henan Province, China.
J Gene Med. 2021 Feb;23(2):e3297. doi: 10.1002/jgm.3297. Epub 2021 Jan 17.
The antibody-dependent enhancement (ADE) of dengue virus (DENV) has critically restricted vaccine development. Prior research suggested pr4 as the probable ADE epitope of DENV.
Chimeric DENV was constructed by replacing the DENV pr4 gene with the corresponding Japanese encephalitis virus (JEV) gene to determine whether it can reduce ADE activities. An alanine scanning method and bioinformatics analysis were utilized to identify the amino acid of pr4 that was crucial as an ADE epitope.
Chimeric virus reduced ADE and virulence. The amino acids at the following locations on the mutant peptides showed significantly reduced binding ability to prM antibody: pr4.5 (position 5 - leucine), pr4.6 (position 6 - leucine), pr4.7 (position 7 - phenyalanine) and pr4.16 (position 16 - cysteine). The four amino acids had formed a pocket-like structure, which could increase the possibility of binding to an antibody.
ADE activities could be reduced by replacing the DENV pr4 gene with the corresponding JEV gene. Leucine at position 5, leucine at position 6, phenyalanine at position 7 and cysteine at position 16 were the key amino acid sites in the ADE response of DENV. The occurrence of ADE can potentially be reduced by the replacement of key amino acids, hence highlighting its possible contribution to dengue vaccine design, paving a way for future vaccine research.
登革热病毒(DENV)的抗体依赖性增强(ADE)极大地限制了疫苗的开发。先前的研究表明 pr4 是 DENV 的可能 ADE 表位。
通过用相应的日本脑炎病毒(JEV)基因替换 DENV pr4 基因来构建嵌合 DENV,以确定它是否可以降低 ADE 活性。采用丙氨酸扫描法和生物信息学分析来鉴定作为 ADE 表位的关键 pr4 氨基酸。
嵌合病毒降低了 ADE 和毒力。突变肽中以下位置的氨基酸显示与 prM 抗体的结合能力显著降低:pr4.5(位置 5-亮氨酸)、pr4.6(位置 6-亮氨酸)、pr4.7(位置 7-苯丙氨酸)和 pr4.16(位置 16-半胱氨酸)。这四个氨基酸形成了一个口袋样结构,增加了与抗体结合的可能性。
用相应的 JEV 基因替换 DENV pr4 基因可降低 ADE 活性。位置 5 的亮氨酸、位置 6 的亮氨酸、位置 7 的苯丙氨酸和位置 16 的半胱氨酸是 DENV ADE 反应中的关键氨基酸位点。通过替换关键氨基酸,可能会降低 ADE 的发生,因此强调了其在登革热疫苗设计中的可能贡献,为未来的疫苗研究铺平了道路。