Pratt William D, Davis Nancy L, Johnston Robert E, Smith Jonathan F
Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702-5011, USA.
Vaccine. 2003 Sep 8;21(25-26):3854-62. doi: 10.1016/s0264-410x(03)00328-1.
The central objective of this research was to test molecularly defined, live attenuated Venezuelan equine encephalitis virus (VEEV) vaccine candidates that were produced through precise genetic manipulation of rationally selected viral nucleotide sequences. Molecular clones of vaccine candidates were constructed by inserting either three independently attenuating mutations or a PE2 cleavage-signal mutation with a second-site resuscitating mutation into full-length cDNA clones. Vaccine candidate viruses were recovered through DNA transcription and RNA transfection of cultured cells, and assessed in rodent and non-human primate models. Based on results from this assessment, one of the PE2 cleavage-signal mutants, V3526, was determined to be the best vaccine candidate for further evaluation for human use.
本研究的核心目标是对通过对合理选择的病毒核苷酸序列进行精确基因操作而产生的分子定义的减毒活委内瑞拉马脑炎病毒（VEEV）候选疫苗进行测试。通过将三个独立的减毒突变或一个带有第二位点复苏突变的PE2裂解信号突变插入全长cDNA克隆中，构建候选疫苗的分子克隆。通过培养细胞的DNA转录和RNA转染回收候选疫苗病毒，并在啮齿动物和非人类灵长类动物模型中进行评估。基于该评估结果，确定PE2裂解信号突变体之一V3526是进一步评估用于人类的最佳候选疫苗。
