Tang Hai-Bo, Lu Zhuan-Ling, Zhong Yi-Zhi, He Xiao-Xia, Zhong Tao-Zhen, Pan Yan, Wei Xian-Kai, Luo Yang, Liao Su-Huan, Minamoto Nobuyuki, Luo Ting Rong
The Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering, State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning, 530004, Guangxi, China.
Virus Genes. 2014 Dec;49(3):417-27. doi: 10.1007/s11262-014-1108-2. Epub 2014 Aug 21.
In this study, a street rabies virus isolate, GXHXN, was obtained from the brain of one rabid cattle in Guangxi province of southern China. To characterize the biological properties of GXHXN, we first evaluated its pathogenicity using 4-week-old adult mice. GXHXN was highly pathogenic with a short incubation period and course of disease. Its LD50 of 10(-6.86)/mL is significantly higher than the LD50 of 10(-5.19)/mL of GXN119, a dog-derived rabies virus isolate. It also displayed a higher neurotropism index than the rRC-HL strain. However, the relative neurotropism index of GXHXN was slightly lower than that of GXN119. Analyzing antigenicity using anti-N and anti-G monoclonal antibodies (MAbs), all tested anti-N MAbs reacted similarly to GXHXN, CVS, and rRC-HL, but the reaction of anti-N MAbs to GXHXN was slightly different from GXN119. Moreover, 2/11 tested anti-G mAbs showed weaker reactivity to GXHXN than rRC-HL, whereas 4/11 showed stronger reactivity to GXHXN than CVS and GXN119, indicating that the structures of G might differ. In order to understand its genetic variation and evolution, the complete GXHXN genome sequence was determined and compared with the known 12 isolates from other mammals. A total of 42 nucleotide substitutions were found in the full-length genome, including 15 non-synonymous mutations. The G gene accounts for the highest nucleotide substitution rate of 0.70 % in ORF and an amino acid substitution rate of 0.95 %. Phylogenetic trees based on the complete genome sequence as well as the N and G gene sequences from 37 known rabies isolates from various mammals demonstrated that the GXHXN is closely related to the BJ2011E isolate from a horse in Beijing, the WH11 isolate from a donkey in Hubei, and isolates from dogs in the Fujian and Zhejiang provinces. These findings will be helpful in exploring the molecular mechanisms underlying interspecies transmission and the genetic variation of the rabies virus in different mammal species.
在本研究中,从中国南方广西省一头患狂犬病的牛脑中获得了一株街毒株狂犬病病毒GXHXN。为了表征GXHXN的生物学特性,我们首先使用4周龄的成年小鼠评估其致病性。GXHXN具有高度致病性,潜伏期和病程较短。其半数致死剂量(LD50)为10(-6.86)/mL,显著高于犬源狂犬病病毒分离株GXN119的LD50(10(-5.19)/mL)。它还表现出比rRC-HL株更高的嗜神经性指数。然而,GXHXN的相对嗜神经性指数略低于GXN119。使用抗N和抗G单克隆抗体(MAbs)分析抗原性,所有测试的抗N单克隆抗体对GXHXN、CVS和rRC-HL的反应相似,但抗N单克隆抗体对GXHXN的反应与GXN119略有不同。此外,11种测试的抗G单克隆抗体中有2种对GXHXN的反应性比rRC-HL弱,而11种中有4种对GXHXN的反应性比CVS和GXN119强,这表明G的结构可能不同。为了了解其遗传变异和进化,测定了GXHXN的完整基因组序列,并与其他哺乳动物的12株已知分离株进行了比较。在全长基因组中总共发现了42个核苷酸替换,包括15个非同义突变。G基因在开放阅读框中的核苷酸替换率最高,为0.70%,氨基酸替换率为0.95%。基于37株来自不同哺乳动物的已知狂犬病分离株的完整基因组序列以及N和G基因序列构建的系统发育树表明,GXHXN与北京一匹马的BJ2011E分离株、湖北一头驴的WH11分离株以及福建和浙江两省的犬分离株密切相关。这些发现将有助于探索狂犬病病毒在不同哺乳动物物种间传播的分子机制及其遗传变异。