H9N2亚型禽流感病毒SD18株的传染性及不同途径感染效果比较

doi:10.16431/j.cnki.1671-7236.2022.04.027

摘要/Abstract

摘要： 【目的】研究H9N2亚型禽流感病毒(Avian influenza virus,AIV)对哺乳动物的传染性和致病性。【方法】采集样品进行病毒的分离鉴定,将分离到的病毒以300 μL/只(10⁷ EID₅₀)的剂量通过滴鼻和肺递送方式感染豚鼠,每组各15只,对2组感染效果进行比较,然后通过分离株在豚鼠中的传播能力试验验证其气溶胶传播能力,通过间接免疫荧光试验检测分离株在人支气管上皮细胞上的复制能力。【结果】试验分离到1株H9N2亚型AIV,命名为SD18,病毒通过肺递送和滴鼻2种攻毒方式感染豚鼠后,肺递送组的排毒量显著高于滴鼻组(P<0.05)。通过对豚鼠肺脏相关模式识别受体和抗病毒蛋白的表达检测发现,2种攻毒方式都能诱导相关的免疫因子Toll样受体3(TLR3)、TLR7、抗黏病毒蛋白(MX)和2',5'-腺苷酸激酶(OAS)表达量极显著上调(P<0.01);2组在攻毒后第6天产生抗体,并呈上升趋势。对SD18 株在豚鼠中的传播能力验证发现,SD18株具有通过直接接触和飞沫传播感染豚鼠的能力,但并未证实气溶胶的产生和传播。对SD18株感染人支气管上皮细胞BEAS-2B后发现,在感染后12 h可检测到病毒,24 h时病毒在细胞内复制能力最强,TLR3、TLR7、MX、OAS、白介素6(IL-6)、IL-8、干扰素-β(IFN-β)相关的免疫因子表达量极显著上调(P<0.01)。【结论】 H9N2亚型AIV SD18株的跨种传播能力变强,具有不经提前适应便可直接感染豚鼠的能力;SD18株具有通过飞沫传播感染豚鼠并使其产生抗体的能力且SD18株可在人支气管上皮细胞BEAS-2B上进行复制。

关键词: H9N2亚型AIV; 肺递送气溶胶感染; 致病性; 飞沫传播

Abstract: 【Objective】 The study was aimed to elucidate the transmission and pathogenicity of H9N2 subtype Avian influenza virus (AIV) in mammals.【Method】 Samples were collected for virus isolation and identification in this study.The isolated virus was used to infect guinea pigs by nasal drops and lung delivery at a dose of 300 μL (10⁷ EID₅₀),with 15 guinea pigs in each group,and the infection effects of the two groups were compared,then the aerosol transmission ability of the isolated strain was verified by the transmission ability experiment in guinea pigs.The replication ability of the isolated strain on human bronchial epithelial cells was detected by indirect immunofluorescence assay.【Result】 A strain of H9N2 subtype AIV was isolated and was named SD18.After guinea pigs were infected with the virus by nasal drip and lung delivery,the guinea pigs infected by lung delivery showed greater virus excretion than the nasal drip (P<0.05).Through the detection the expression of pattern recognition receptors and antiviral proteins in lungs of guinea pigs,the expression of immunologic factors Toll-like receptor 3 (TLR3),TLR7,myxovirus-resistance (MX),2'-5'oligoadenylates synthesis (OAS) had extremely significantly increased in both modes of infection (P<0.01).Antibodies of H9N2 subtype AIV began to be developed on the 6th day and showed an upward trend.The transmission ability of SD18 strain in guinea pigs was verified.It was found that SD18 strain had the ability to infect guinea pigs through direct contact and droplet transmission,but the generation and transmission of aerosols had not been confirmed.When human bronchial epithelial BEAS-2B cells were infected with SD18 strain,the virus could be detected at 12 h after infection,its replicability within cells peaked at 24 h,and the expression of immune factors TLR3,TLR7,MX,OAS,interleukin 6 (IL-6),IL-8 and interferon β(IFN-β) were extremely significantly upregulated (P<0.01).【Conclusion】 The interspecies transmission of SD18 strain had considerably become stronger,it could directly infect guinea pigs without passage adaptation.SD18 strain had the ability to infect guinea pigs and produce antibodies through droplets,and SD18 strain could replicate on human bronchial epithelial BEAS-2B cells.

Key words: H9N2 subtype AIV; lung-delivered aerosol infection; pathogenicity; droplet transmission

中图分类号:

S852.65^＋7

侯晓红, 王蒙, 武博, 龙宪荣, 赵孝民, 高静, 戴培强, 柴同杰. H9N2亚型禽流感病毒SD18株的传染性及不同途径感染效果比较[J]. 中国畜牧兽医, 2022, 49(4): 1449-1460.

HOU Xiaohong, WANG Meng, WU Bo, LONG Xianrong, ZHAO Xiaomin, GAO Jing, DAI Peiqiang, CHAI Tongjie. Infectivity of H9N2 Subtype Avian Influenza Virus SD18 Strain and Comparison of the Effects of Different Routes of Infection[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1449-1460.

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