Saxena Sonal, Dahiya Shyam S, Sonwane Arvind A, Patel Chhabi Lal, Saini Mohini, Rai A, Gupta Praveen K
Division of Animal Biotechnology, Indian Veterinary Research Institute, Izatnagar 243122, India.
Vaccine. 2008 Dec 2;26(51):6592-601. doi: 10.1016/j.vaccine.2008.09.055.
A sindbis virus replicon-based DNA vaccine encoding rabies virus glycoprotein (G) was developed by subcloning rabies G gene into a sindbis virus replicon-based vaccine vector (pAlpha). The self-amplification of RNA transcripts and translation efficiency of rabies G was analyzed in pAlpha-Rab-G-transfected mammalian cells using RT-PCR, SDS-PAGE and Western blot analysis. The transfected cells also showed induction of apoptosis which is an important event in the enhancement of immune responses. Further, immune responses induced with replicon-based rabies DNA vaccine (pAlpha-Rab-G) was compared with conventional rabies DNA vaccine and commercial cell culture vaccine (Rabipur) in intramuscularly injected mice. The mice immunized with replicon-based rabies DNA vaccine induced humoral and cell mediated immune responses better than conventional rabies DNA vaccine however, comparable to Rabipur vaccine. On challenge with rabies virus CVS strain, replicon-based rabies DNA vaccine conferred complete protection similar to Rabipur. These results demonstrate that replicon-based rabies DNA vaccine is effective in inducing both humoral and cellular immune responses and can be considered as effective vaccine against rabies.
通过将狂犬病病毒糖蛋白（G）基因亚克隆到基于辛德毕斯病毒复制子的疫苗载体（pAlpha）中，构建了一种编码狂犬病病毒糖蛋白的基于辛德毕斯病毒复制子的DNA疫苗。利用逆转录聚合酶链反应（RT-PCR）、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质免疫印迹分析，在pAlpha-Rab-G转染的哺乳动物细胞中分析了狂犬病病毒G基因RNA转录本的自我扩增和翻译效率。转染的细胞还表现出凋亡诱导，这是增强免疫反应中的一个重要事件。此外，在肌肉注射的小鼠中，比较了基于复制子的狂犬病DNA疫苗（pAlpha-Rab-G）与传统狂犬病DNA疫苗和商业细胞培养疫苗（Rabipur）诱导的免疫反应。用基于复制子的狂犬病DNA疫苗免疫的小鼠诱导的体液免疫和细胞介导免疫反应优于传统狂犬病DNA疫苗，但与Rabipur疫苗相当。在用狂犬病病毒CVS株攻击时，基于复制子的狂犬病DNA疫苗提供了与Rabipur相似的完全保护。这些结果表明，基于复制子的狂犬病DNA疫苗在诱导体液免疫和细胞免疫反应方面是有效的，可被视为一种有效的狂犬病疫苗。
