水禽H6N6亚型禽流感病毒在猪和人的肺组织中的复制能力的体外研究  

Replication capacity of aquatic bird-originated H6N6 subtype of avian influenza virus in lung tissues of swine and human:an in vitro study

作　　者：林裕贵 庞夏凤 李程颐 孙增辉[1] 朱银川[1] 王熹婧 林春秀 张增峰[1] LIN Yu-gui;PANG Xia-feng;LI Cheng-yi;SUN Zeng-hui;ZHU Yin-chuan;WANG Xi-jing;LIN Chun-xiu;ZHANG Zeng-feng(Department of Microbiology,Guangxi Medical University,Nanning 530021,China;School of Food Science and Technology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]广西医科大学微生物学教研室,南宁市530021 [2]江南大学食品学院,江苏省无锡市214122

出　　处：《广西医学》2021年第8期962-965,971,共5页Guangxi Medical Journal

基　　金：国家自然科学基金(31660041);广西自然科学基金(2018GXNSFAA281300);广西研究生教育创新计划(YCSW2020124);广西本级财政科技计划(2018JJA140657)。

摘　　要：目的研究水禽H6N6亚型禽流感病毒(AIV)在体外猪和人的肺组织中的复制能力。方法选取两株分离自鸭或鹅的H6N6亚型AIV(A/DK/GX/750/2010和A/GS/GX/399/2010),行基因测序和遗传进化分析了解其基因来源。将每株病毒分别体外接种于猪肺和人肺组织,接种后24 h、48 h、72 h收集组织,取组织匀浆液和培养上清液在10 d鸡胚中分离培养病毒,并行苏木精-伊红染色观察组织病理改变,采用免疫组化法检测病毒核蛋白的表达。结果基因进化分析显示H6N6亚型AIV A/DK/GX/750/2010、A/GS/GX/399/2010属于欧亚谱系的Group-Ⅱ(ST2853-like),为同一重配病毒的不同变型。两株H6N6亚型AIV体外接种于猪肺和人肺组织后,组织匀浆液和培养上清液中均可分离到病毒,免疫组化检测提示病毒核蛋白呈阳性,其主要表达在肺泡细胞和肺泡巨噬细胞,苏木精-伊红染色显示肺组织内出现肺泡细胞坏死和肺泡破坏,人肺组织还伴有炎性细胞浸润。结论水禽H6N6亚型AIV能在体外猪和人的肺组织中有效地复制,提示该病毒具有感染哺乳动物猪及人类的潜能,必须密切监测其流行及进化演变情况,以防止其感染人类。Objective To investigate the replication capacity of aquatic bird-originated H6N6 subtype of avian influenza virus(AIV)in lung tissues of swine and human in vitro.Methods Two strains of H6N6 subtype AIV(A/DK/GX/750/2010 and A/GS/GX/399/2010)isolated from duck or goose were obtained,and gene sequencing and phylogenetic analysis were conducted to investigate the genome origin of the viruses.The viruses were inoculated into lung tissues of swine and human in vitro,and the tissues were collected 24,48 and 72 hours after inoculation with tissue homogenate and culture supernatants for viral isolation and culture in 10-day chicken embryo,histopathological changes were observed by hematoxylin-eosin staining,and the expression of viral nucleoprotein was detected by immunohistochemistry assay.Results Phylogenetic analysis revealed that H6N6 subtype AIV A/DK/GX/750/2010 and A/GS/GX/399/2010 belonged to Group-Ⅱ(ST2853-like)of Eurasian lineage and different types of the same reassortment virus.After two strains of H6N6 subtype AIV inoculated into the lung tissues of swine and human,the viruses could be isolated from tissue homogenate or culture supernatants,viral nucleoprotein was confirmed as positive by immunohistochemistry assay and was mainly expressed in alveolar cells and macrophages,and hematoxylin-eosin staining showed that alveolar cell necrosis and alveolar cavity destruction in lung tissues and inflammatory cell infiltration in human lung tissues were observed.Conclusion Aquatic bird-originated H6N6 subtype AIV can replicate effectively in swine and human lung tissues in vitro,which indicates that the virus has the potential to infect mammals swine and human.It is necessary to monitor closely the epidemic and evolution of aquatic bird-originated H6N6 subtype AIV in case of infection to human.

关 键 词：禽流感病毒 水禽 H6N6亚型 复制能力 猪 人 体外实验 肺组织 

分 类 号：R373[医药卫生—病原生物学]