| In recent yearsthe unpredictable infection of the avian influenza virus could trigger a global pandemic. The virus has been a hot research topic in the world.The increasing or decreasing of haemagglutinin (HA) glycosylation sites ofthe avian influenza virus combined with the glycan structure changes, may change the virus antigenicity, change different bonding specificity and strengthentheforce between the viruses and the different genus host receptors. Besides HA, the glycosylation of another glycoprotein-neuraminidase (NA) in influenza virus particle surface can adjust their own enzyme activity, can influence their hydrolytic release, sialic acid ability and can prevent itself from being digested by the host protease. This is why the NA can affect the virulence of virus and specificity of the hosts. First, the HA and NA are synthesized in the endoplasmic reticulum in host cells. Then, they use the host cell of sugar chain to synthesis the system so it cancarry on the N-glycosylation modification in the endoplasmic reticulum and the Golgi body. However, there are few known avian influenza virus subtypes strains from a poultry source and a humanized cell proliferation, after its protein glycosylation changes (including glycosylation sites occupied and glycan structure difference). Therefore, it is necessary to do the further research.The Purpose of this experiment is to use lectin microarray technology for comparative analyze of the different host proliferative H9N2subtype and theglycan spectrum of glycoprotein. The experiment can be divided into three parts:(1) establishing the model of using human embryo lung cells (MRC-5) to proliferate the avian influenza virus;(2) using this model proliferate H9N2avian influenza virus subtypes, and through the lectin microarray technology, perform a comparative analyzeof H9N2avian influenza virus subtypes from MRC-5proliferation and its negative contrast glycoprotein glycan spectrum;(3) using lectinmicroarray technology to analyze our laboratory saved H9N2avian influenza virus subtypesfrom a chicken embryo (CE), and perform a comparative analyze of the twodifferent virus subtypes’ glycan spectrum from MRC-5and CE respectively to get the different glycan spectrum.The experimental results show that,the abundance of the glycan, which can be recognized by lectins HHL, VVA, NPL, UEA-I, PWM, and SNAare significantly higher in the virus from MRC-5than the virus from CE; Otherwise, the abundance of the glycan, which can be recognized by lectins ECA, WFA, MAL-Ⅱ, PHA-E, AAL, GSL-I, LCA, RCA120, STL, Con-A, DSA, PSA, and ACA are obviously higherin the virus from CE than the virus from MRC-5. According to these distinctive recognition glycanof different lectins, the analysis results are as follows:(1) H9N2subtype avian influenza virus,which proliferated from MRC-5contains moreSia2-6Galβ1-4Glc(NAc)than from CE.(2) H9N2subtype avian influenza virus from CE contains moreSia2-3Gal(31-4Glc(NAc), but not present in MRC-5.(3) The abundance of Mannose(Man)in H9N2avian influenza virus subtypes from MRC-5is much higher thanfromCE.From MRC-5, the most structure of Man is Non-substituteda-1,6Man, but from CE the structure of H9N2avian influenza virus subtypes of Man is branched and terminal mannose.(4)The abundance of Fucose and Fucoseα-1,6GlcNAc(core fucose) are quite highin H9N2subtype avianinfluenza virus from CE. The abundance of Fucoseal-2Galβ1-4Glc(NAc) from MRC-5is higher than from CE.(5) The abundance of Gal or GalNAc in H9N2avian influenza virus subtypes proliferated from CEis significantly higher than fromMRC-5.Through the comparative analysis of the different glycan chain spectrum in H9N2avian influenza virus from the MRC-5cell lines and chicken embryo. We know much more about the different glycan spectrum characteristics of H9N2avian influenza virus from both humanized and poultry source. After knowing that will contribute to clearify the molecular mechanism of avian influenza virus subtype crossing the sepecies in mammals in further research. |
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