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Science china(life Sciences) 2009年第6期52卷 545-550页

作者： li WeiZhong,wang gefei,zeng jun,ZHANG DanGui,ZHANG Heng,CHEN XiaoXuan,CHEN YouYing & li kangsheng department of microbiology and immunology,shantou university medical college,shantou 515041,china 1. department of microbiology and immunology Shantou University Medical College Shantou 515041 China

Influenza A virus NS1 protein is an important regulatory factor with multiple functions and contributes greatly to viral *** the present study,transcription-activating potential of NS1 from different influenza A virus subtypes was examined in yeast two-hybrid *** bait vectors contain-ing different NS1 genes,along with an empty prey vector,were transformed into yeast AH109(for growth assay on QDO plate and α-galactosidase assay),and Y187(for β-galactosidase assay).AH109 transformants with NS1 gene from H1N1,H5N1,and H9N2 viruses grew vigorously on the QDO plate and secreted high level of α-***,Y187 bearing the above NS1 genes exhibited en-hanced β-galactosidase ***,H3N2-NS1-transformed AH109 and Y187 yeasts did not grow on QDO plate and secrete β-galactosidase,*** findings denote the remarkable variation in NS1 proteins from different influenza A virus subtypes on the transcription-stimulating capability in yeast.

关键词： Influenza virus NS1 protein yeast two-hybridation transcription-activation

Effects of NS1 variants of H5N1 influenza virus on interferon induction, TNFa response and p53 activity

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Cellular & Molecular immunology 2010年第3期7卷 235-242页

作者： Weizhong li gefei wang Heng Zhang Gang Xin Dangui Zhang jun zeng Xiaoxuan Chen Yanxuan Xu Youhong Cui kangsheng li department of microbiology and immunology Key Immunopathology Laboratory of Guangdong ProvinceShantou University Medical CollegeShantouChina

Non-structural protein 1(NS1)is an important virulence factor of the highly pathogenic H5N1 avian influenza virus.A five-amino-acid(5 aa)deletion at position 80–84 and an aspartic acid to glutamic acid substitution at position 92(D92E)are two major NS1 mutations that are highly correlated with enhanced *** investigate the effect of these mutations in H5N1 virulence,three H5N1-NS1 variants were constructed:NS51(lacking 5 aa at position 80–84),NS51(I)(carrying a 5-aa insertion at position 80–84)and NS51(IM)(carrying both the 5-aa insertion and the D92E mutation).We examined the effects of these mutations on interferon(IFN)induction,tumor-necrosis factor(TNF)a response,p53 activity and *** found that the D92E mutation eliminated NS1’s repressive effect on IFN induction,while the 5-aa deletion resulted in enhanced resistance to TNFa *** also observed that all three variants exhibited a similar suppressive effect on p53 transcriptional activity,although none of them significantly influenced apoptosis of host *** findings shed new light on the role of NS1 in the pathogenicity of H5N1 virus.

关键词： H5N1 influenza virus interferon NS1 variants p53 TNFa

A型流感病毒NS1蛋白羧基端PL结构域影响NS1在细胞核内的定位

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生物化学与生物物理进展 2010年第9期37卷 975-982页

作者：张丹桂李卫中王革非张衡曾俊陈幼莹张驰曾祥兴李康生汕头大学医学院微生物学及免疫学教研室广东省高校免疫病理学重点实验室汕头515031

A型流感病毒NS1蛋白羧基端4个氨基酸可以与PDZ结构域(the domain of PSD95,Dig and ZO-1)相结合,称为PL结构域(PDZ ligand domain).对不同亚型或毒株的流感病毒而言,其NS1蛋白PL结构域的组成存在比较大的差异.有研究发现这种差异能够影... 详细信息

A型流感病毒NS1蛋白羧基端4个氨基酸可以与PDZ结构域(the domain of PSD95,Dig and ZO-1)相结合,称为PL结构域(PDZ ligand domain).对不同亚型或毒株的流感病毒而言,其NS1蛋白PL结构域的组成存在比较大的差异.有研究发现这种差异能够影响NS1与宿主细胞蛋白的相互作用进而影响病毒的致病力.为进一步探讨PL结构域对NS1蛋白生物学特性的影响,首先构建出4种不同亚型流感病毒(H1N1、H3N2、H5N1、H9N2)来源的NS1绿色荧光蛋白表达质粒.在此基础上,对野生型H3N2病毒NS1表达质粒进行人工改造,将其PL结构域缺失或者替换为其他亚型流感病毒的PL结构域,制备出4种重组NS1蛋白表达质粒.通过比较上述不同NS1蛋白在HeLa细胞中的定位情况发现,只有野生型H3N2病毒的NS1蛋白可以定位于核仁当中,而野生型H1N1、H5N1、H9N2病毒的NS1蛋白以及PL结构域缺失或替代的H3N2病毒NS1蛋白都不能定位于核仁.而通过比较上述NS1蛋白在流感病毒易感的MDCK细胞中的定位,进一步发现所有这些蛋白均不定位于核仁.上述结果表明:PL结构域的不同可以明显影响NS1蛋白在HeLa细胞核内的定位和分布,这有可能造成其生物学功能的差异.同时,NS1蛋白在细胞核内的定位还与宿主细胞的来源有着密切关系.

关键词： A型流感病毒 NS1蛋白 PL结构域核仁

Heterologous interactions between NS1 proteins from different influenza A virus subtypes/strains

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Science china(life Sciences) 2012年第6期55卷 507-515页

作者： li WeiZhong ZHANG Heng wang gefei ZHANG Chi zeng XiangXing liU Hui CHEN XiaoXuan XU YanXuan li kangsheng department of microbiology and immunology Key Immunopathology Laboratory of Guangdong ProvinceShantou University Medical CollegeShantou 515041China department of Veterinary Medicine University of MarylandCollege ParkMD 20742USA department of Ultrasonic Central Hospital of Luoyang CityLuoyang 471009China

Non-structural protein 1 （NS1） of the influenza virus plays a crucial role in modulating the host immune response and facili- tating virus replication. The formation of a homodimer or an oligomer is necessary for NSI to exert its function efficiently. In the present study, the NS 1 protein from the A/shantou/602/06（H3N2） virus （herein abbreviated as NS32） was found to interact with NS1 from A/shantou/169/O6（H1N1）, A/Chicken/Guangdong/1/05（HSN1） and A/Quail/Hong Kong/G1/97（H9N2）（abbre- viated as NS11, NS51 and NS92, respectively） viruses, although NS32 shares 17.4%-20.9% sequence diversity with NS11, NS51 and NS92. This indicates that the heterologous interactions between NS1 proteins from different influenza A virus sub- types/strains may be a common event during co-infection.

关键词： influenza A virus NS1 protein heterologous interaction

Coxsackieviruses Infection Induced Cytopathy and Cytokine Secretion in Astrocyte

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Coxsackieviruses Infection Induced Cytopathy and Cytokine Se...

Apoptosis and Proinflammatory Cytokine Responses of Primary Mouse Microglia and Astrocytes Induced by Human H1N1 and Avian H5N1 Influenza Viruses

2012年鄂粤微生物学学术年会——湖北省暨武汉微生物学会成立六十年庆祝大会

作者： zeng jun wang gefei li Weizhong Chen Xiaoxuan li kangsheng department of microbiology and immunology Shantou University Medical College

Coxsackieviruses most commonly infect children,causing mild symptoms,but on occasion cause severe meningitis,encephalitis in the *** investigate the mechanisms of such inflammatory diseases,virus replication,the morpholgical alterations,potential inflammation cytokine and chemokine gene activation in primary mouse astrocyes were investigated following infection with coxsackievirus

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Cellular & Molecular immunology 2008年第2期5卷 113-120页

作者： gefei wang Juan Zhang Weizhong li Yun Su Yuanli Gao Heng Zhang Guimei lin Xiaoyang Jiao kangsheng li department of microbiology and immunology Shantou University MedicalCollege Shantou 515041 China The Key Immunopathology Laboratory of Guangdong Province Shantou University Medical College Shantou 515041 China

Patients with an influenza virus infection can be complicated by acute encephalopathy and encephalitis. To investigate the immune reactions involved in the neurocomplication, mouse microglia and astrocytes were isolated, infected with human H1N1 and avian H5N1 influenza viruses, and examined for their immune responses. We observed homogeneously distributed viral receptors, sialic acid （SA）-a2,3-Galactose （Gal） and SA-a2,6-Gal, on microglia and astrocytes. Both viruses were replicative and productive in microglia and astrocytes. Virus-induced apoptosis and cytopathy in infected cells were observed at 24 h post-infection （p.i.）. Expression of IL-1β, IL-6 and TNF-a mRNA examined at 6 h and 24 h p.i. was up-regulated, and their expression levels were considerably higher in H5N1 infection. The amounts of secreted proinflammatory IL-1β, IL-6 and TNF-a at 6 h and 24 h p.i. were also induced, with greater induction by H5N1 infection. This study is the first demonstration that both human H1N1 and avian H5N1 influenza viruses can infect mouse microglia and astrocytes and induce apoptosis, cytopathy, and proinflammatory cytokine production in them in vitro. Our results suggest that the direct cellular damage and the consequences of immunopathological injury in the CNS contribute to the influenza viral pathogenesis. Cellular ＆ Molecular immunology.

关键词： microglia astrocyte apoptosis cytokine influenza virus

基于免疫信息学的SARS-CoV-2表位筛选及疫苗分子设计分析

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中国免疫学杂志 2022年第7期38卷 838-847页

作者：邓辉雄王革非(指导) 李雁嫘宋鑫利谷李铭李蕊(指导) 汕头大学医学院微生物学与免疫学教研室病原与免疫学研究中心汕头515041

目的:基于新型冠状病毒(SARS-CoV-2)全病毒或Spike蛋白作为免疫原可能存在的无关抗原表位干扰,筛选鉴定优势保护性抗原表位,构建候选多价表位疫苗。方法:以免疫信息学确定诱导主要中和抗体、激活细胞免疫应答的细胞保护性抗原表位,以Rap... 详细信息

目的:基于新型冠状病毒(SARS-CoV-2)全病毒或Spike蛋白作为免疫原可能存在的无关抗原表位干扰,筛选鉴定优势保护性抗原表位,构建候选多价表位疫苗。方法:以免疫信息学确定诱导主要中和抗体、激活细胞免疫应答的细胞保护性抗原表位,以Raptor X、trRosetta预测蛋白疫苗的二级、三级结构,并对候选蛋白疫苗进行二硫键的设计及对接分析,以C-ImmSim服务器模拟预测分析多表位疫苗的免疫原性和免疫应答。结果:实验筛选了Spike蛋白免疫原性强的B细胞表位,具有高保守性和人群覆盖度广的IFN-γ、IL-4阳性的Th表位及CTL表位,计算模拟验证了疫苗的免疫应答特性。结论:信息学分析结果初步显示候选表位疫苗具有良好的平衡体液免疫和细胞免疫应答能力。

关键词：新冠病毒 Spike蛋白抗原表位平衡免疫免疫信息学

血清CHO、HDL、LDL、sdLDL、SOD与冠心病的相关性研究

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国际检验医学杂志 2019年第15期40卷 1814-1816页

作者：吴愿如黄华曾彩凤邝紫华王革非蔡蓁汕头大学医学院微生物与免疫学教研室广东汕头515041 深圳市龙岗区第二人民医院检验科广东深圳518112 深圳市孙逸仙心血管医院广东深圳518000

目的评价血清胆固醇(CHO)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)、小而密低密度脂蛋白(sdLDL)及超氧化物歧化酶(SOD)与冠心病的相关性。方法选取冠心病患者131例作为冠心病组,116例健康体检者作为对照组,采用全自动生化分析仪检测各... 详细信息

目的评价血清胆固醇(CHO)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL)、小而密低密度脂蛋白(sdLDL)及超氧化物歧化酶(SOD)与冠心病的相关性。方法选取冠心病患者131例作为冠心病组,116例健康体检者作为对照组,采用全自动生化分析仪检测各组血清CHO、HDL、LDL、sdLDL、SOD水平,并对检验结果进行统计学处理。结果冠心病组血清CHO、LDL及sdLDL水平高于对照组,差异有统计学意义(P<0.05)。HDL及SOD水平低于对照组,差异有统计学意义(P<0.05)。sdLDL与CHO、LDL均呈正相关(r分别为0.517、0.651,P<0.005),sdLDL与HDL、SOD均呈负相关(r分别为-0.238、-0.395,P<0.05),多因素logistic回归分析显示,sdLDL是冠心病的独立危险因素。结论冠心病患者血清sdLDL水平升高,SOD水平降低,sdLDL为冠心病的独立危险因素。

关键词：小而密低密度脂蛋白超氧化物歧化酶冠心病