Generation and characterization of P gene-deficient rabies virus.
Generation and characterization of P gene-deficient rabies virus.
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DOI：
10.1016/j.virol.2003.10.001
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发表时间：
2004-01
期刊：
影响因子：
3.7
通讯作者：
Y. Shoji;S. Inoue;K. Nakamichi;I. Kurane;T. Sakai;K. Morimoto
中科院分区：
文献类型：
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作者：
Y. Shoji;S. Inoue;K. Nakamichi;I. Kurane;T. Sakai;K. Morimoto
P基因缺失的狂犬病病毒(RV)是从缺乏全P基因的Hep-Flury株的cDNA中反向遗传而来的。通过拯救病毒，使用与缺陷基因功能互补的细胞系，对缺陷病毒进行繁殖和扩增。P基因缺陷(def-P)病毒复制其基因组，并在构成表达P蛋白的细胞系中产生后代病毒，尽管它的生长速度与亲本株相比略有减慢。相反，当Def-P病毒感染不表达P蛋白的细胞时，感染宿主中不会产生子代病毒。然而，我们发现Def-P病毒具有在感染宿主中执行初级转录(通过病毒粒子相关聚合酶)的能力，而不需要重新合成P蛋白。DEF-P病毒在成年和哺乳小鼠中是无致病性的，即使在脑内接种也是如此。在小鼠体内接种DEF-P病毒可诱导高水平的病毒中和抗体(VNA)，并对致死性狂犬病感染产生保护性免疫。这些结果证明了基因缺陷病毒作为一种新型狂犬病减毒活疫苗的潜在用途。
Rabies virus (RV) deficient in the P gene was generated by reverse genetics from cDNA of HEP-Flury strain lacking the entire P gene. The defective virus was propagated and amplified by rescue of virus, using a cell line that complemented the functions of the deficient gene. The P gene-deficient (def-P) virus replicated its genome and produced progeny viruses in the cell lines that constitutively expressed the P protein, although it grew at a slightly retarded rate compared to the parental strain. In contrast, no progeny virus was produced in the infected host when the def-P virus-infected cells that did not express the P protein. However, we found that the def-P virus had the ability to perform primary transcription (by the virion-associated polymerase) in the infected host without de novo P protein synthesis. The def-P virus was apathogenic in adult and suckling mice, even when inoculated intracranially. Inoculation of def-P virus in mice induced high levels of virus-neutralizing antibody (VNA) and conferred protective immunity against a lethal rabies infection. These results demonstrate the potential utility of gene-deficient virus as a novel live attenuated rabies vaccine.