It remains challenging to develop animal models of lung infection and severe pneumonia by severe acute respiratory syndrome coronavirus(SARS-CoV) and Middle East respiratory syndrome cornavirus(MERS-Co V) without high...It remains challenging to develop animal models of lung infection and severe pneumonia by severe acute respiratory syndrome coronavirus(SARS-CoV) and Middle East respiratory syndrome cornavirus(MERS-Co V) without high level of containment. This inevitably hinders understanding of virushost interaction and development of appropriate countermeasures. Here we report that intranasal inoculation of sublethal doses of murine coronavirus mouse hepatitis virus A-59(MHV-A59), a hepatic and neuronal tropic coronavirus, can induce acute pneumonia and severe lung injuries in C57BL/6 mice. Inflammatory leukocyte infiltrations, hemorrhages and fibrosis of alveolar walls can be observed 2-11 days after MHV-A59 infection. This pathological manifestation is associated with dramatical elevation of tissue IP-10 and IFN-γ and moderate increase of TNF-α and IL-1β, but inability of anti-viral type I interferon response. These results suggest that intranasal infection of MHV-A59 would serve as a surrogate mouse model of acute respiratory distress syndrome by SARS-CoV and MERS-CoV infections.展开更多
Coronaviruses (CoVs) are generally associated with respiratory and enteric infections and have long been recognized as important pathogens of livestock and companion animals. Mouse hepatitis virus (MHV) is a widely st...Coronaviruses (CoVs) are generally associated with respiratory and enteric infections and have long been recognized as important pathogens of livestock and companion animals. Mouse hepatitis virus (MHV) is a widely studied model system for Coronavirus replication and pathogenesis. In this study,we created a MHV-A59 temperature sensitive (ts) mutant Wu"-ts18(cd) using the recombinant vaccinia reverse genetics system. Virus replication assay in 17C1-1 cells showed the plaque phenotype and replication characterization of constructed Wu"-ts18(cd) were indistinguishable from the reported ts mutant Wu"-ts18. Then we cultured the ts mutant Wu"-ts18(cd) at non-permissive temperature 39.5°C,which "forced" the ts recombinant virus to use second-site mutation to revert from a ts to a non-ts phenotype. Sequence analysis showed most of the revertants had the same single amino acid mutation at Nsp16 position 43. The single amino acid mutation at Nsp16 position 76 or position 130 could also revert the ts mutant Wu"-ts18 (cd) to non-ts phenotype,an additional independent mutation in Nsp13 position 115 played an important role on plaque size. The results provided us with genetic information on the functional determinants of Nsp16. This allowed us to build up a more reasonable model of CoVs replication-transcription complex.展开更多
Infections by coronaviruses such as severe acute respiratory syndrome （SARS） coronavirus （SCoV） and mouse hepatitis virus A59 （MHV-A59） result in very little type I interferon （IFN） production by host cells, w...Infections by coronaviruses such as severe acute respiratory syndrome （SARS） coronavirus （SCoV） and mouse hepatitis virus A59 （MHV-A59） result in very little type I interferon （IFN） production by host cells, which is potentially responsible for the rapid viral growth and severe immunopathology associated with SARS. However, the molecular mechanisms for the low IFN production in cells infected with coronaviruses remain unclear. Here, we provide evidence that Papain-like protease domain 2 （PLP2）, a catalytic domain of the nonstructural protein 3 （nsp3） of MHV-A59, can bind to IRF3, cause its deubiquitination and prevent its nuclear translocation. As a consequence, co-expression of PLP2 strongly inhibits CARDIF-, TBK1- and IRF3-mediated IFNp reporter activities. In addition, we show that wild-type PLP2 but not the mutant PLP2 lacking the deubiquitinase （DUB） activity can reduce IFN induction and promote viral growth in cells infected with VSV. Thus, our study uncovered a viral DUB which coronaviruses may use to escape from the host innate antiviral responses.展开更多
目的了解我国华北、东北、华东、中南、西南和西北地区,2006年1～59岁人群乙型肝炎(乙肝)病毒(Hepatitis B Virus,HBV)感染现状及乙肝疫苗(Hepatitis B Vaccine,HepB)接种情况。方法系统分析2006年全国乙肝血清流行病学调查资料。结果...目的了解我国华北、东北、华东、中南、西南和西北地区,2006年1～59岁人群乙型肝炎(乙肝)病毒(Hepatitis B Virus,HBV)感染现状及乙肝疫苗(Hepatitis B Vaccine,HepB)接种情况。方法系统分析2006年全国乙肝血清流行病学调查资料。结果华北、东北、华东、中南、西南和西北地区,2006年1～59岁人群HepB接种率分别为50.81%、53.78%、50.02%、47.93%、31.21%和47.60%;乙肝病毒表面抗原(HBV Surface Antigen,HBsAg)流行率分别3.83%、5.56%、7.76%、9.12%、7.92%、4.87%;抗乙肝病毒表面抗原抗体(Antibody to HBsAg,Anti-HBs)流行率分别42.79%、46.44%、48.76%、57.36%、49.74%、46.42%。结论我国六个区域2006年1～59岁人群HBsAg流行率均有不同程度下降,Anti-HBs流行率有所上升,我国自1992年HepB纳入儿童计划免疫管理效果显著。展开更多
基金supported in part by grants from the Natural Science Foundation of China (30430640,31030031)the National Basic Research Program of MOST (2004BA519A61,2003CB514116,2006CB504300)Natural Science Foundation of China (31400765)
文摘It remains challenging to develop animal models of lung infection and severe pneumonia by severe acute respiratory syndrome coronavirus(SARS-CoV) and Middle East respiratory syndrome cornavirus(MERS-Co V) without high level of containment. This inevitably hinders understanding of virushost interaction and development of appropriate countermeasures. Here we report that intranasal inoculation of sublethal doses of murine coronavirus mouse hepatitis virus A-59(MHV-A59), a hepatic and neuronal tropic coronavirus, can induce acute pneumonia and severe lung injuries in C57BL/6 mice. Inflammatory leukocyte infiltrations, hemorrhages and fibrosis of alveolar walls can be observed 2-11 days after MHV-A59 infection. This pathological manifestation is associated with dramatical elevation of tissue IP-10 and IFN-γ and moderate increase of TNF-α and IL-1β, but inability of anti-viral type I interferon response. These results suggest that intranasal infection of MHV-A59 would serve as a surrogate mouse model of acute respiratory distress syndrome by SARS-CoV and MERS-CoV infections.
基金Research Grants from State Key Laboratory of Pathogen and Biosecurity (SKLPBS0918)
文摘Coronaviruses (CoVs) are generally associated with respiratory and enteric infections and have long been recognized as important pathogens of livestock and companion animals. Mouse hepatitis virus (MHV) is a widely studied model system for Coronavirus replication and pathogenesis. In this study,we created a MHV-A59 temperature sensitive (ts) mutant Wu"-ts18(cd) using the recombinant vaccinia reverse genetics system. Virus replication assay in 17C1-1 cells showed the plaque phenotype and replication characterization of constructed Wu"-ts18(cd) were indistinguishable from the reported ts mutant Wu"-ts18. Then we cultured the ts mutant Wu"-ts18(cd) at non-permissive temperature 39.5°C,which "forced" the ts recombinant virus to use second-site mutation to revert from a ts to a non-ts phenotype. Sequence analysis showed most of the revertants had the same single amino acid mutation at Nsp16 position 43. The single amino acid mutation at Nsp16 position 76 or position 130 could also revert the ts mutant Wu"-ts18 (cd) to non-ts phenotype,an additional independent mutation in Nsp13 position 115 played an important role on plaque size. The results provided us with genetic information on the functional determinants of Nsp16. This allowed us to build up a more reasonable model of CoVs replication-transcription complex.
基金These authors contributed equally to this work. We thank Drs S Vaidya and E Chow （University of California Los Angeles, USA） for their help in setting up critical experimental systems. We greatly thank Dr K Holmes （University of Colorado Health Sciences Center, USA） for sharing with us 17C1-1 cell line and helping to optimize the protocol to produce high titered MHV-A59 virus stock. We also thank Drs R Baric and L Su （University of North Carolina, USA） for the gift of MHV-A59 and guidance of virus infection. We thank Dr K Lim （National Neuroscience Institute, Singapore） for the gift of Ubi plasmids. We thank Dr M Wathelet （University of Cincinnati College of Medicine, USA） for sharing the nsp3 construct. Also we thank Dr G Gao （Institute of Biophysics, CAS） for providing us with VSV. This research was partly supported by grants from the National Natural Science Foundation of China （30728006） to Genhong Cheng and the National Basic Research Program of MOST （2004BA519A61, 2006CB504300, 2007DFC30190） to Hong Tang.
文摘Infections by coronaviruses such as severe acute respiratory syndrome （SARS） coronavirus （SCoV） and mouse hepatitis virus A59 （MHV-A59） result in very little type I interferon （IFN） production by host cells, which is potentially responsible for the rapid viral growth and severe immunopathology associated with SARS. However, the molecular mechanisms for the low IFN production in cells infected with coronaviruses remain unclear. Here, we provide evidence that Papain-like protease domain 2 （PLP2）, a catalytic domain of the nonstructural protein 3 （nsp3） of MHV-A59, can bind to IRF3, cause its deubiquitination and prevent its nuclear translocation. As a consequence, co-expression of PLP2 strongly inhibits CARDIF-, TBK1- and IRF3-mediated IFNp reporter activities. In addition, we show that wild-type PLP2 but not the mutant PLP2 lacking the deubiquitinase （DUB） activity can reduce IFN induction and promote viral growth in cells infected with VSV. Thus, our study uncovered a viral DUB which coronaviruses may use to escape from the host innate antiviral responses.
文摘目的了解我国华北、东北、华东、中南、西南和西北地区,2006年1～59岁人群乙型肝炎(乙肝)病毒(Hepatitis B Virus,HBV)感染现状及乙肝疫苗(Hepatitis B Vaccine,HepB)接种情况。方法系统分析2006年全国乙肝血清流行病学调查资料。结果华北、东北、华东、中南、西南和西北地区,2006年1～59岁人群HepB接种率分别为50.81%、53.78%、50.02%、47.93%、31.21%和47.60%;乙肝病毒表面抗原(HBV Surface Antigen,HBsAg)流行率分别3.83%、5.56%、7.76%、9.12%、7.92%、4.87%;抗乙肝病毒表面抗原抗体(Antibody to HBsAg,Anti-HBs)流行率分别42.79%、46.44%、48.76%、57.36%、49.74%、46.42%。结论我国六个区域2006年1～59岁人群HBsAg流行率均有不同程度下降,Anti-HBs流行率有所上升,我国自1992年HepB纳入儿童计划免疫管理效果显著。
