[1]赵国洪,王继华,李华春,等.口蹄疫兔化弱毒疫苗ZB株L蛋白N2D、M143L和E147G氨基酸突变对病毒毒力影响的研究[J].中国预防兽医学报,2018,(09):822-827.[doi:0.3969/j.issn.1008-0589.201712043]
 ZHAO Guo-hong,WANG Ji-hua,LI Hua-chun,et al.Foot-and-mouth disease virus of attenuated vaccine ZB strain L protein mutations N2D, M143L and E147G do not impair viral virulence[J].Chinese journal of preventive veterinary medicine,2018,(09):822-827.[doi:0.3969/j.issn.1008-0589.201712043]
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口蹄疫兔化弱毒疫苗ZB株L蛋白N2D、M143L和E147G氨基酸突变对病毒毒力影响的研究()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2018年09
页码:
822-827
栏目:
免疫学
出版日期:
2018-09-17

文章信息/Info

Title:
Foot-and-mouth disease virus of attenuated vaccine ZB strain L protein mutations N2D, M143L and E147G do not impair viral virulence
文章编号:
1008-0589(2018)9-0822-06
作者:
 

赵国洪12王继华3李华春12何于雯12朱明旺3*信爱国12*

 (1. 云南省畜牧兽医科学院 国家口蹄疫专业实验室(昆明),云南 昆明 650224;2. 云南省畜牧兽医科学院 云南省热带亚热带动物病毒病重点实验室,云南 昆明 650224;3. 云南省兽医生物制品研制中心,云南 保山 678000)
Author(s):
 

ZHAO Guo-hong12 WANG Ji-hua3 LI Hua-chun12 HE Yu-wen12ZHU Ming-wang3* XIN Ai-guo12*

(1. National Professional Laboratory of Foot-and-mouth disease (Kunming), Yunnan Animal Science and Veterinary Institute, Kunming 650224,China; 2. Yunnan Tropical and Subtropical Animal Virus Disease Laboratory, Yunnan Animal Science and Veterinary Institute,
Kunming 650224, China; 3. Yunnan Provincial Research Center for Veterinary Biological Products, Baoshan 678000, China)
关键词:
口蹄疫病毒兔化弱毒疫苗株ZB株前导蛋白点突变
Keywords:
foot-and-mouth disease virus  rabbit-attenuated vaccine strain  ZB strain  leader protein  point mutation
分类号:
S852.65
DOI:
0.3969/j.issn.1008-0589.201712043
文献标志码:
A
摘要:
:口蹄疫病毒(FMDV)前导蛋白Lpro是病毒的毒力因子之一。前期研究发现,FMDV兔化弱毒疫苗ZB株传代减毒致弱后,前导蛋白Lpro内仅存在3个氨基酸点突变(N2D、M143L和E147G)。为确定这3个突变对病毒生物学特性的影响,本研究利用弱毒疫苗ZB株感染性分子克隆为骨架,构建获得含有这3个氨基酸回复突变的重组病毒rZBatt-L,并比较其与亲本病毒rZBatt生物学特性。结果显示,rZBatt-L和rZBatt在BHK-21细胞中均可以产生细胞病变效应,形成大小相似的噬斑,而在牛原代肾细胞(BK)中均不产生CPE;二者在BHK-21细胞中的生长曲线与病毒RNA复制动态相似(p>0.05),对乳鼠的致病性也无显著差异;测定了rZBatt-L和rZBatt分别感染BK诱导I型干扰素(IFN-α 和β)能力,结果显示rZBatt-L和rZBatt均可以在感染早期诱导IFN- α表达,而感染晚期抑制IFN- α表达。相对的是,rZBatt-L和rZBatt感染细胞后均可以诱导IFN-β含量增加,并且二者诱导IFN-β含量无差异(p>0.05)。结果表明,FMDV弱毒疫苗ZB株Lpro蛋白中的N2D、M143L和E147G突变不改变病毒的生物学特性,对病毒毒力不产生明显的影响。本研究明确了Lpro蛋白突变不是ZB株致弱的关键氨基酸位点。
 
Abstract:
Foot-and-mouth disease virus (FMDV) leader proteinase (Lpro) is one of the viral virulence factors. Previously, the only three amino acid mutations (N2D, M143L and E147G) had occurred in Lpro of FMDV attenuated vaccine ZB strain during the attenuation process. In this study, a chimeric virus (rZBatt-L) which encoded the Lpro of virulent ZB virus containing the N2D, M143L and E147G mutations was rescued. Comparison of rZBatt-L with its parental plasmid-derived rescued virus (rZBatt) demonstrated that these two viruses displayed similar plaque morphologies in BHK-21 cells, but no plaque formation was observed in primary fetal bovine kidney (BK) cells. Analysis of TCID50 growth curves and FMDV RNA levels revealed the similar replication kinetics between rZBatt-L and rZBatt (p>0.05). The pathogenicity in suckling mice of these two viruses showed no significant difference (p>0.05). Furthermore, the levels of type I interferon (IFN-α and IFN-β) in rZBatt-L or rZBatt virus-infected BK were detected. The production of IFN-α in BK were elevated in early-phase infection, but it was inhibited in the late infection phase. In contrast, the IFN-β levels were increased in BK infected with either rZBatt-L or rZBatt, without any significant difference (p>0.05). The rZBatt-L displayed similar characteristics as its parental virus rZBatt. These results indicated that the mutations in the Lpro protein of ZB attenuated vaccine virus were not to be major determinants of the attenuation phenotype.

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(本文编辑:李   爽)

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更新日期/Last Update: 2018-10-19