古典H1N1亚型猪流感病毒PB2 K627E突变对病毒复制能力的影响

doi:10.11843/j.issn.0366-6964.2016.03.017

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (3): 543-548.doi: 10.11843/j.issn.0366-6964.2016.03.017

古典H1N1亚型猪流感病毒PB2 K627E突变对病毒复制能力的影响

汪秀会¹，宫晓倩¹，阮宝阳^1，2，刘晓敏¹，汪琪¹，张鹏^1，2，李泽君¹，周艳君¹，童武¹，郑浩¹，童光志^1*，于海^1*

（1.中国农业科学院上海兽医研究所，上海 200241；2.山东农业大学动物科技学院，泰安 271000）

收稿日期:2015-08-04 出版日期:2016-03-23 发布日期:2016-03-21
通讯作者: 童光志，研究员，E-mail：gztong@shvri.ac.cn；于海，副研究员，E-mail：haiyu@shvri.ac.cn
作者简介:汪秀会（1988- ），女，山东聊城人，硕士生，主要从事猪流感病毒分子致病机制的研究，E-mail：wangxiuhui123@163.com
基金资助:
国家青年自然科学基金（31201916）；上海市自然科学基金青年项目（12ZR1453500）；中央级公益性科研院所基本科研业务费项目（2015JB07）；中国农业科学院创新工程科研团队“动物流感病毒病原生态学”项目

The Effect of PB2 K627E Mutation on the Replication Competence of Classical H1N1 Subtype Swine Influenza Virus

WANG Xiu-hui¹，GONG Xiao-qian¹，RUAN Bao-yang^1，2，LIU Xiao-min¹，WANG Qi¹，ZHANG Peng^1，2，LI Ze-jun¹，ZHOU Yan-jun¹，TONG Wu¹，ZHENG Hao¹，TONG Guang-zhi^1* ，YU Hai^1*

（1.Shanghai Veterinary Research Institute，Chinese Academy of Agricultural Sciences，Shanghai 200241，China；2.College of Veterinary Medicine，Shandong Agricultural University，Tai’an 271000，China）

Received:2015-08-04 Online:2016-03-23 Published:2016-03-21

摘要/Abstract

摘要：

旨在探索古典H1N1亚型猪流感病毒A/Swine/Guangdong/1/2011(H1N1) PB2 K627E突变对病毒复制能力的影响。利用反向遗传操作技术和点突变技术成功获得古典H1N1亚型猪流感病毒的拯救株（627K）和突变株（627E）后，分别在MDCK细胞和小鼠体内对拯救株和突变株的复制能力进行比较。体外MDCK细胞试验结果显示，突变株与拯救株相比，细胞病变(CPE)减小、空斑形态减小、生长曲线差异极其显著，表明突变株在MDCK细胞上的复制能力减弱。小鼠攻毒试验结果表明，突变株对小鼠体重变化的影响不明显，在小鼠肺中的病毒滴度明显下降，引起病理损伤较轻。体外MDCK细胞及小鼠攻毒试验结果均表明，突变株与拯救株相比复制能力降低。该结果不仅证实了猪流感病毒 PB2 627位氨基酸是一种重要的毒力分子标记，同时还为进一步阐明其致病机制提供了条件。

关键词: 古典H1N1亚型猪流感病毒, PB2 K627E突变, 体外MDCK细胞试验, 小鼠攻毒试验, 复制能力

Abstract:

This experiment was conducted to explore the effect of PB2 K627E mutation on the replication competence of classical H1N1 subtype swine influenza virus(A/Swine/Guangdong/1/2011).The rescued and mutant strains of classical H1N1 subtype swine influenza virus were obtained by reverse genetics and point mutation technology，and researches were taken on MDCK cells and mice.The results on MDCK cells showed that，compared with the rescued strain，the mutant virus had a slight degree of CPE；the morphology of plaque of mutant virus was much smaller；the growth curve showed that it had a significant distinct difference.The results of experiments in mice showed that the mutant strain had a low pathogenicity and could not lead the mice loss weight，and virus titer in lung tissues on the 3rd and 5th day post challenge showed that there was a significant distinct difference between the two strains and the pathological damage caused by mutant one was not serious.The results on MDCK and mice both showed that the mutant strain’s replication competence was lower than the rescued one.This conclusion demonstrates that the amino acid of PB2 627 is not only one of the important virulence markers，but also provided the condition to further clarify its pathogenic mechanism.

Key words: classical H1N1 subtype swine influenza, PB2 K627E mutation, vitro experiment in MDCK cells, vivo experiment in mice, virus replication

中图分类号:

S852.659.5

汪秀会，宫晓倩，阮宝阳，刘晓敏，汪琪，张鹏，李泽君，周艳君，童武，郑浩，童光志，于海. 古典H1N1亚型猪流感病毒PB2 K627E突变对病毒复制能力的影响[J]. 畜牧兽医学报, 2016, 47(3): 543-548.

WANG Xiu-hui，GONG Xiao-qian，RUAN Bao-yang，LIU Xiao-min，WANG Qi，ZHANG Peng，LI Ze-jun，ZHOU Yan-jun，TONG Wu，ZHENG Hao，TONG Guang-zhi，YU Hai. The Effect of PB2 K627E Mutation on the Replication Competence of Classical H1N1 Subtype Swine Influenza Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(3): 543-548.

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古典H1N1亚型猪流感病毒PB2 K627E突变对病毒复制能力的影响

The Effect of PB2 K627E Mutation on the Replication Competence of Classical H1N1 Subtype Swine Influenza Virus

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