Human respiratory syncytial virus（RSV） is an important pediatric pathogen of lower respiratory tract worldwide. No vaccines and antiviral drugs are available. Herein the use of an RNA polymerase I-driven RSV minigen...Human respiratory syncytial virus（RSV） is an important pediatric pathogen of lower respiratory tract worldwide. No vaccines and antiviral drugs are available. Herein the use of an RNA polymerase I-driven RSV minigenome for analyzing RSV replication and screening anti-RSV drugs was investigated. The RNA polymerase I（Pol I） was used to transcribe RSV minigenome from the constructed plasmid, designated p HM-RSV-Gluc, of minigenome c DNA which comprised trailer region, gene start sequence（GS）, reverse complementary copy of Gaussia luciferase（Gluc） gene, gene end sequence（GE）, and leader region in the direction of 5＇–3＇end and was flanked by promoter and terminator of Pol I. The expression of Gluc was confirmed in p HM-RSV-Gluc transfected HEp-2 cells following RSV infection and had the characteristics of dose-dependent, which provided a rapid, sensitive, and quantitative method for quantifying virus titers and screening antiviral drugs.展开更多
The current scenario of COVID-19 makes us to think about the devastating diseases that kill so many people every year.Analysis of viral proteins contributes many things that are utterly useful in the evolution of ther...The current scenario of COVID-19 makes us to think about the devastating diseases that kill so many people every year.Analysis of viral proteins contributes many things that are utterly useful in the evolution of therapeutic drugs and vaccines.In this study,sequence and structure of fusion glycoproteins and major surface glycoproteins of respiratory syncytial virus(RSV)were analysed to reveal the stability and transmission rate.RSV A has the highest abundance of aromatic residues.The Kyte–Doolittle scale indicates the hydrophilic nature of RSV A protein which leads to the higher transmission rate of this virus.Intra-protein interactions such as carbonyl interactions,cation-pi,and salt bridges were shown to be greater in RSV A compared to RSV B,which might lead to improved stability.This study discovered the presence of a network aromatic–sulphur interaction in viral proteins.Analysis of ligand binding pocket of RSV proteins indicated that drugs are performing better on RSV B than RSV A.It was also shown that increasing the number of tunnels in RSV A proteins boosts catalytic activity.This study will be helpful in drug discovery and vaccine development.展开更多
目的:通过研究金欣口服液含药血清对呼吸道合胞病毒(RSV)活化诱导的Toll样受体3(TLR3)的干预作用,探讨其治疗RSV肺炎可能的免疫学机制。方法:RSV感染体外培养的RAW264.7细胞,采用金欣口服液含药血清进行干预,并设利巴韦林阳性对照,24h...目的:通过研究金欣口服液含药血清对呼吸道合胞病毒(RSV)活化诱导的Toll样受体3(TLR3)的干预作用,探讨其治疗RSV肺炎可能的免疫学机制。方法:RSV感染体外培养的RAW264.7细胞,采用金欣口服液含药血清进行干预,并设利巴韦林阳性对照,24h后收集细胞,Real time RT-PCR法测定TLR3 mRNA的表达变化;激光共聚焦显微镜观察免疫荧光细胞化学染色法处理的各组细胞TLR3表达水平;ELISA法检测各组细胞培养上清中白细胞介素-6(IL-6)含量,在核酸及蛋白水平探讨金欣口服液含药血清抗RSV感染的作用机制。结果:RSV感染RAW264.7细胞后24h,细胞中TLR3 mRNA和蛋白表达水平以及细胞培养上清中IL-6表达明显升高(P<0.01),而金欣组TLR3及IL-6表达均显著低于RSV感染组(P<0.01,P<0.05),且效果优于利巴韦林组(P<0.01,P<0.05)。结论:金欣口服液含药血清能下调RSV活化诱导的TLR3及其下游炎症因子IL-6的高表达,推测其为金欣口服液抗RSV感染的机制之一。展开更多
基金supported by National Major Scientific and Technological Special Project for ‘‘AIDS and Viral Hepatitis and Other Major Infectious Diseases Prevention and Control’’ during the Twelfth Five-year Plan Period (No. 2013ZX10004-601)
文摘Human respiratory syncytial virus（RSV） is an important pediatric pathogen of lower respiratory tract worldwide. No vaccines and antiviral drugs are available. Herein the use of an RNA polymerase I-driven RSV minigenome for analyzing RSV replication and screening anti-RSV drugs was investigated. The RNA polymerase I（Pol I） was used to transcribe RSV minigenome from the constructed plasmid, designated p HM-RSV-Gluc, of minigenome c DNA which comprised trailer region, gene start sequence（GS）, reverse complementary copy of Gaussia luciferase（Gluc） gene, gene end sequence（GE）, and leader region in the direction of 5＇–3＇end and was flanked by promoter and terminator of Pol I. The expression of Gluc was confirmed in p HM-RSV-Gluc transfected HEp-2 cells following RSV infection and had the characteristics of dose-dependent, which provided a rapid, sensitive, and quantitative method for quantifying virus titers and screening antiviral drugs.
文摘The current scenario of COVID-19 makes us to think about the devastating diseases that kill so many people every year.Analysis of viral proteins contributes many things that are utterly useful in the evolution of therapeutic drugs and vaccines.In this study,sequence and structure of fusion glycoproteins and major surface glycoproteins of respiratory syncytial virus(RSV)were analysed to reveal the stability and transmission rate.RSV A has the highest abundance of aromatic residues.The Kyte–Doolittle scale indicates the hydrophilic nature of RSV A protein which leads to the higher transmission rate of this virus.Intra-protein interactions such as carbonyl interactions,cation-pi,and salt bridges were shown to be greater in RSV A compared to RSV B,which might lead to improved stability.This study discovered the presence of a network aromatic–sulphur interaction in viral proteins.Analysis of ligand binding pocket of RSV proteins indicated that drugs are performing better on RSV B than RSV A.It was also shown that increasing the number of tunnels in RSV A proteins boosts catalytic activity.This study will be helpful in drug discovery and vaccine development.
文摘目的:通过研究金欣口服液含药血清对呼吸道合胞病毒(RSV)活化诱导的Toll样受体3(TLR3)的干预作用,探讨其治疗RSV肺炎可能的免疫学机制。方法:RSV感染体外培养的RAW264.7细胞,采用金欣口服液含药血清进行干预,并设利巴韦林阳性对照,24h后收集细胞,Real time RT-PCR法测定TLR3 mRNA的表达变化;激光共聚焦显微镜观察免疫荧光细胞化学染色法处理的各组细胞TLR3表达水平;ELISA法检测各组细胞培养上清中白细胞介素-6(IL-6)含量,在核酸及蛋白水平探讨金欣口服液含药血清抗RSV感染的作用机制。结果:RSV感染RAW264.7细胞后24h,细胞中TLR3 mRNA和蛋白表达水平以及细胞培养上清中IL-6表达明显升高(P<0.01),而金欣组TLR3及IL-6表达均显著低于RSV感染组(P<0.01,P<0.05),且效果优于利巴韦林组(P<0.01,P<0.05)。结论:金欣口服液含药血清能下调RSV活化诱导的TLR3及其下游炎症因子IL-6的高表达,推测其为金欣口服液抗RSV感染的机制之一。
