Human adenoviruses(HAdVs)are highly contagious pathogens with various genotypes implicated in acute respiratory disease(ARD)and linked to fatality,especially in immunosuppressed patients,young children,and military re...Human adenoviruses(HAdVs)are highly contagious pathogens with various genotypes implicated in acute respiratory disease(ARD)and linked to fatality,especially in immunosuppressed patients,young children,and military recruits.Currently,no vaccines or specific drugs are approved for clinical use.The hosts of adenoviruses are strictly species-specific,which strongly limits the development of vaccines and drugs against HAdVs.In this study,immunocompetent BALB/c mice were challenged with different doses of human adenovirus type 5(HAdV-5)via tail intravenous injection(i.v.).All mice challenged with a high dose of HAdV-5(3.21010 TCID50/kg)died within 3–5 days,while those receiving a low dose of HAdV-5(8109 or 4109 TCID50/kg)survived.Interestingly,among the mice receiving a medium dose of HAdV-5(1.61010 TCID50/kg),60%(n¼3/5)of male mice died,while all female mice survived.This suggests that male mice may be more susceptible to HAdV-5 infection than female mice,consistent with clinical findings in children.HAdV-5 DNA was mainly distributed in the liver,followed by the spleen and lung.Pathological changes were observed in the lung,liver,and spleen,with severity increasing in correlation with the virus challenge dosage.Transcriptome and qPCR analyses of the liver indicated that the down-regulated expression of the H2-Aa,H2-Ea-ps,CD74,and H2-Eb1 genes in male mice,as well as the AHR gene in female mice,may contribute to the observed higher mortality rates in male mice.Therefore,this effective,feasible,and costefficient mouse model could serve as a candidate for evaluating HAdV vaccines and anti-adenovirus therapeutics.展开更多
Human adenoviruses(HAd Vs)are highly contagious and result in large number of acute respiratory disease(ARD)cases with severe morbidity and mortality.Human adenovirus type 3(HAd V-3)is the most common type that causes...Human adenoviruses(HAd Vs)are highly contagious and result in large number of acute respiratory disease(ARD)cases with severe morbidity and mortality.Human adenovirus type 3(HAd V-3)is the most common type that causes ARD outbreaks in Asia,Europe,and the Americas.However,there is currently no vaccine approved for its general use.The hexon protein contains the main neutralizing epitopes,provoking strong and lasting immunogenicity.In this study,a novel recombinant and attenuated adenovirus vaccine candidate against HAd V-3 was constructed based on a commercially-available replication-defective HAd V-5 gene therapy and vaccine vector.The entire HAd V-3 hexon gene was integrated into the E1 region of the vector by homologous recombination using a bacterial system.The resultant recombinants expressing the HAd V-3 hexon protein were rescued in AD293 cells,identified and characterized by RT-PCR,Western blots,indirect immunofluorescence,and electron microscopy.This potential vaccine candidate had a similar replicative efficacy as the wild-type HAd V-3 strain.However,and importantly,the vaccine strain had been rendered replication-defective and was incapable of replication in A549 cells after more than twentygeneration passages in AD293 cells.This represents a significant safety feature.The mice immunized both intranasally and intramuscularly by this vaccine candidate raised significant neutralizing antibodies against HAd V-3.Therefore,this recombinant,attenuated,and safe adenovirus vaccine is a promising HAd V-3 vaccine candidate.The strategy of using a clinically approved and replication-defective HAd V-5 vector provides a novel approach to develop universal adenovirus vaccine candidates against all the other types of adenoviruses causing ARDs and perhaps other adenovirus-associated diseases.展开更多
Restriction endonuclease analysis(REA),or restriction fragment length polymorphism(RFLP),was useful for identifying and determining the relatedness and putative identities of microbial strains(Tang et al.,1997)and for...Restriction endonuclease analysis(REA),or restriction fragment length polymorphism(RFLP),was useful for identifying and determining the relatedness and putative identities of microbial strains(Tang et al.,1997)and for characterizing and discriminating large numbers of samples inexpensively in the past。展开更多
The current pandemic of COVID-19 is fueled by more infectious emergent Omicron variants.Ongoing concerns of emergent variants include possible recombinants,as genome recombination is an important evolutionary mechanis...The current pandemic of COVID-19 is fueled by more infectious emergent Omicron variants.Ongoing concerns of emergent variants include possible recombinants,as genome recombination is an important evolutionary mechanism for the emergence and re-emergence of human viral pathogens.In this study,we identified diverse recombination events between two Omicron major subvariants(BA.1 and BA.2)and other variants of concern(VOCs)and variants of interest(VOIs),suggesting that co-infection and subsequent genome recombination play important roles in the ongoing evolution of SARS-CoV-2.Through scanning high-quality completed Omicron spike gene sequences.展开更多
基金supported by grants from the National Natural Science Foundation of China(32170139 and 92269103)the R&D Program of Guangzhou Laboratory(No.SRPG22-006)+4 种基金the Guangdong Basic and Applied Basic Research Foundation(2022A1515011190)the Science and Technology Program of Guangzhou,China(202201011115)the Open Foundation of Key Laboratory of Viral Pathogenesis&Infection Prevention and Control(Jinan University),Ministry of Education(2023VPPC-R07)China Postdoctoral Science Foundation(2023M731321)the Lifting Project Foundation of the Affiliated Guangdong Second Provincial General Hospital of Jinan University(TJGC-2022004).
文摘Human adenoviruses(HAdVs)are highly contagious pathogens with various genotypes implicated in acute respiratory disease(ARD)and linked to fatality,especially in immunosuppressed patients,young children,and military recruits.Currently,no vaccines or specific drugs are approved for clinical use.The hosts of adenoviruses are strictly species-specific,which strongly limits the development of vaccines and drugs against HAdVs.In this study,immunocompetent BALB/c mice were challenged with different doses of human adenovirus type 5(HAdV-5)via tail intravenous injection(i.v.).All mice challenged with a high dose of HAdV-5(3.21010 TCID50/kg)died within 3–5 days,while those receiving a low dose of HAdV-5(8109 or 4109 TCID50/kg)survived.Interestingly,among the mice receiving a medium dose of HAdV-5(1.61010 TCID50/kg),60%(n¼3/5)of male mice died,while all female mice survived.This suggests that male mice may be more susceptible to HAdV-5 infection than female mice,consistent with clinical findings in children.HAdV-5 DNA was mainly distributed in the liver,followed by the spleen and lung.Pathological changes were observed in the lung,liver,and spleen,with severity increasing in correlation with the virus challenge dosage.Transcriptome and qPCR analyses of the liver indicated that the down-regulated expression of the H2-Aa,H2-Ea-ps,CD74,and H2-Eb1 genes in male mice,as well as the AHR gene in female mice,may contribute to the observed higher mortality rates in male mice.Therefore,this effective,feasible,and costefficient mouse model could serve as a candidate for evaluating HAdV vaccines and anti-adenovirus therapeutics.
基金supported by Grants from the National Key Research and Development Program of China(2018YFE0204503)National Natural Science Foundation of China(31570155,31370199)+1 种基金Natural Science Foundation of Guangdong Province(2018B030312010)the Guangzhou Healthcare Collaborative Innovation Major Project(201803040004,201803040007)。
文摘Human adenoviruses(HAd Vs)are highly contagious and result in large number of acute respiratory disease(ARD)cases with severe morbidity and mortality.Human adenovirus type 3(HAd V-3)is the most common type that causes ARD outbreaks in Asia,Europe,and the Americas.However,there is currently no vaccine approved for its general use.The hexon protein contains the main neutralizing epitopes,provoking strong and lasting immunogenicity.In this study,a novel recombinant and attenuated adenovirus vaccine candidate against HAd V-3 was constructed based on a commercially-available replication-defective HAd V-5 gene therapy and vaccine vector.The entire HAd V-3 hexon gene was integrated into the E1 region of the vector by homologous recombination using a bacterial system.The resultant recombinants expressing the HAd V-3 hexon protein were rescued in AD293 cells,identified and characterized by RT-PCR,Western blots,indirect immunofluorescence,and electron microscopy.This potential vaccine candidate had a similar replicative efficacy as the wild-type HAd V-3 strain.However,and importantly,the vaccine strain had been rendered replication-defective and was incapable of replication in A549 cells after more than twentygeneration passages in AD293 cells.This represents a significant safety feature.The mice immunized both intranasally and intramuscularly by this vaccine candidate raised significant neutralizing antibodies against HAd V-3.Therefore,this recombinant,attenuated,and safe adenovirus vaccine is a promising HAd V-3 vaccine candidate.The strategy of using a clinically approved and replication-defective HAd V-5 vector provides a novel approach to develop universal adenovirus vaccine candidates against all the other types of adenoviruses causing ARDs and perhaps other adenovirus-associated diseases.
基金supported by the National Natural Science Foundation of China (31570155 and 31370199)"Young Top-notch Talents" of the Guangdong Province Special Support Program (2014)+3 种基金the Excellent Young Teacher Training Plan of Guangdong Province (Yq2013039)the Guangzhou Healthcare Collaborative Innovation Major Project (201400000002)funded by the China Scholarship Council (CSC No. 201508440056) as a Visiting Scholar (2015-2016)supported by a summer research grant to D.S. from the Office of the Vice President for Research at George Mason University
文摘Restriction endonuclease analysis(REA),or restriction fragment length polymorphism(RFLP),was useful for identifying and determining the relatedness and putative identities of microbial strains(Tang et al.,1997)and for characterizing and discriminating large numbers of samples inexpensively in the past。
基金We gratefully acknowledge the authors,originating and submitting laboratories of the sequences from GISAID’s EpiCoV™Database that this study used.This work was supported by grants from the National Key Research and Development Program of China(2018YFE0204503)National Natural Science Foundation of China(32170139 and 81730061)+2 种基金Natural Science Foundation of Guangdong Province(2018B030312010,2021A1515010788,and 2022A1515011190)the Fundamental Research Funds for the Central Universities(21622101)Guangdong Science and Technology Program key projects(2021B1212030014).
文摘The current pandemic of COVID-19 is fueled by more infectious emergent Omicron variants.Ongoing concerns of emergent variants include possible recombinants,as genome recombination is an important evolutionary mechanism for the emergence and re-emergence of human viral pathogens.In this study,we identified diverse recombination events between two Omicron major subvariants(BA.1 and BA.2)and other variants of concern(VOCs)and variants of interest(VOIs),suggesting that co-infection and subsequent genome recombination play important roles in the ongoing evolution of SARS-CoV-2.Through scanning high-quality completed Omicron spike gene sequences.
