Objective Recombination events are common and serve as the primary driving force of diverse human adenovirus(HAdV),particularly in children with acute respiratory tract infections(ARIs).Therefore,continual monitoring ...Objective Recombination events are common and serve as the primary driving force of diverse human adenovirus(HAdV),particularly in children with acute respiratory tract infections(ARIs).Therefore,continual monitoring of these events is essential for effective viral surveillance and control.Methods Respiratory specimens were collected from children with ARIs between January 2022 and December 2023.The penton base,hexon,and fiber genes were amplified from HAdV-positive specimens and sequenced to determine the virus type.In cases with inconsistent typing results,genes were cloned into the pGEM-T vector to detect recombination events.Metagenomic next-generation sequencing(mNGS)was performed to characterize the recombinant HAdV genomes.Results Among 6,771 specimens,277(4.09%,277/6,771)were positvie for HAdV,of which 157(56.68%,157/277)were successfully typed,with HAdV-B3 being the dominant type(91.08%,143/157),and 14(5.05%,14/277)exhibited inconsistent typing results,six of which belonged to species B.The penton base genes of these six specimens were classified as HAdV-B7,whereas their hexon and fiber genes were classified as HAdV-B3,resulting in a recombinant genotype designated P7H3F3,which closely resembled HAdV-B114.Additionally,a partial gene encoding L152/55 kD was identified,which originated from HAdV-B16.Conclusion A novel recombinant,P7H3F3,was identified,containing sequences derived from HAdV-B3 and HAdV-B7,which is similar to HAdV-B114,along with additional sequences from HAdV-B16.展开更多
Human adenovirus type 108(HAdV-108)has been detected in multiple countries,including China,and is associated with severe acute respiratory infection(ARI)in children,with reported fatalities.However,studies on HAdV-108...Human adenovirus type 108(HAdV-108)has been detected in multiple countries,including China,and is associated with severe acute respiratory infection(ARI)in children,with reported fatalities.However,studies on HAdV-108 remain limited.This study aimed to investigate the clinical and genetic characteristics of HAdV-108 in ARI children in China.From 2014 to 2024,6720 respiratory samples were collected from hospitalized children with ARI at ten hospitals across northern and southern China,of which 505(7.51%)tested positive for HAdV.The whole-genome and three major capsid protein genes were amplified and sequenced for bioinformatics analysis,which revealed that among 317 HAdV-isolated samples,21(6.62%)were identified as HAdV-108,ranking third after HAdV-114 and HAdV-7.Clinical analysis of HAdV-108-positive cases showed that the main manifestations were cough and fever.Seven children had gastrointestinal symptoms,and two children without underlying diseases were diagnosed with severe pneumonia.Phylogenetic analysis of wholegenome sequences revealed distinct predominant epidemic branches between domestic and international strains,with one strain obtained in this study forming an independent branch.Hexon protein exhibited the fastest evolution rate,lowest identity,and greatest amino acid variability,while fiber protein displayed the slowest evolution rate,highest identity,and greatest conservation and stability.Compared with the earliest reported HAdV-108 strain,three amino acid deletions were identified in the RGD loop region of penton base protein,resulting in potential structural change.Recombination analysis identified five distinct recombination patterns.In vitro experiments demonstrated that HAdV-108 had proliferation capacity comparable to other species C adenoviruses.In summary,HAdV-108 has persistently circulated in China,causing severe ARIs and concurrent gastrointestinal manifestations.Cluster3 was the predominant epidemic branch in China.HAdV-108 exhibited significant intratype genetic variation,with random and diverse recombination events.展开更多
During 2018–2019,a severe human adenovirus(HAdV)infection outbreak occurred in southern China.Here,we screened 18 respiratory pathogens in 1704 children(≤14 years old)hospitalized with acute respiratory illness in G...During 2018–2019,a severe human adenovirus(HAdV)infection outbreak occurred in southern China.Here,we screened 18 respiratory pathogens in 1704 children(≤14 years old)hospitalized with acute respiratory illness in Guangzhou,China,in 2019.In total,151 patients had positive HAdV test results;34.4%(52/151)of them exhibited severe illness.HAdV infection occurred throughout the year,with a peak in summer.The median patient age was 3.0(interquartile range:1.1–5.0)years.Patients with severe HAdV infection exhibited increases in12 clinical indexes(P≤0.019)and decreases in four indexes(P≤0.007),compared with patients exhibiting nonsevere infection.No significant differences were found in age or sex distribution according to HAdV infection severity(P>0.05);however,the distributions of comorbid disease and HAdV co-infection differed according to HAdV infection severity(P<0.05).The main epidemic types were HAdV-3(47.0%,71/151)and HAdV-7(46.4%,70/151).However,the severe illness rate was significantly higher in patients with HAdV-7(51.4%)than in patients with HAdV-3(19.7%)and other types of HAdV(20%)(P<0.001).Sequencing analysis of genomes/capsid genes of 13 HAdV-7 isolates revealed high similarity to previous Chinese isolates.A representative HAdV-7isolate exhibited a similar proliferation curve to the curve described for the epidemic HAdV-3 strain Guangzhou01(accession no.DQ099432)(P>0.05);the HAdV-7 isolate exhibited stronger virulence and infectivity,compared with HAdV-3(P<0.001).Overall,comorbid disease,HAdV co-infection,and high virulence and infectivity of HAdV-7 were critical risk factors for severe HAdV infection;these data can facilitate treatment,control,and prevention of HAdV infection.展开更多
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(HAdVs) commonly cause many diseases such as respiratory diseases, gastroenteritis, cystitis worldwide. HAdV-3,-7,-4 and emergent HAdV-55 and HAdV-14 are the most important types causing severe respi...Human adenoviruses(HAdVs) commonly cause many diseases such as respiratory diseases, gastroenteritis, cystitis worldwide. HAdV-3,-7,-4 and emergent HAdV-55 and HAdV-14 are the most important types causing severe respiratory diseases. There is no effective drug available for clinical treatment, and no vaccine available for the general population.Therefore, it is important to investigate the seroprevalence against HAdV for developing novel vaccines and vectors. In this study, we investigated the seroprevalence and titer levels of neutralizing antibodies(NAb) against HAdV-3,-4,-7,-14,-55,and-11 in total 278 healthy populations between 0 months and 49 years of age(228 children and 50 adults) from Guangzhou. In children under the age of 18 years, the seropositive rates were significantly increased against HAdV-3 at12.07%, 33.96%, and 64.29% and against HAdV-7 at 0%, 18.87%, and 19.05% in age groups of 1–2, 3–5, and 6–17 years,respectively. The seroprevalence was very low(0% - 8.1%) for all other four types. In adults aged between 18 and49 years, HAdV-3,-4, and-7(> 50.00%) were the most common types, followed by HAdV-14(38.00%),-55(34.00%),and-11(24.00%). Adults tended to have high NAb titers against HAdV-4 and-55. HAdV-55-seropositive donors tended to be HAdV-11-and HAdV-14-seropositive. These results indicated the low level of herd immunity against all six HAdV types in young children, and HAdV-14,-55,-11 in adults from Guangzhou City. Our findings demonstrate the importance of monitoring HAdV types and developing vaccines against HAdV for children and adults.展开更多
Human adenovirus type 55(HAdV-B55) is a re-emergent acute respiratory disease pathogen that causes adult communityacquired pneumonia(CAP). Previous studies have shown that the receptor of HAdV-B14, which genome is hig...Human adenovirus type 55(HAdV-B55) is a re-emergent acute respiratory disease pathogen that causes adult communityacquired pneumonia(CAP). Previous studies have shown that the receptor of HAdV-B14, which genome is highly similar with HAdV-B55, is human Desmoglein 2(DSG2). However, whether the receptor of HAdV-B55 is DSG2 is undetermined because there are three amino acid mutations in the fiber gene between HAdV-B14 and HAdV-B55. Here, firstly we found the 3T3 cells, a mouse embryo fibroblast rodent cell line which does not express human DSG2, were able to be infected by HAdV-B55 after transfected with pcDNA3.1-DSG2, while normal 3T3 cells were still unsusceptible to HAdV-B55 infection. Next, A549 cells with h DSG2 knock-down by siRNA were hard to be infected by HAdV-B3/-B14/-B55, while the control siRNA group was still able to be infected by all these types of HAdVs. Finally, immunofluorescence confocal microscopy indicated visually that Cy3-conjugated HAdV-B55 viruses entered A549 cells by binding to DSG2 protein.Therefore, DSG2 is a major receptor of HAdV-B55 causing adult CAP. Our finding is important for better understanding of interactions between adenoviruses and host cells and may shed light on the development of new drugs that can interfere with these processes as well as for the development of potent prophylactic vaccines.展开更多
Nosocomial infections are common in pediatric patients and can be fatal in infants and immunocompromised patients. In September 2018, a high positive rate of human adenovirus HAdV was occurred among hospitalized child...Nosocomial infections are common in pediatric patients and can be fatal in infants and immunocompromised patients. In September 2018, a high positive rate of human adenovirus HAdV was occurred among hospitalized children in the Children's Hospital Affiliated to the Capital Institute of Paediatrics in Beijing. To investigate whether this outbreak of HAdV was related to nosocomial infections or the result of community infections, we collected respiratory specimens from patients with acute respiratory infections in a respiratory ward during June to December 2018, and screened for respiratory viruses. Among 1,840 cases included, 95(5.2%, 95/1840) were positive for HAdV and 81 were genotyped based on phylogenetic analysis, including seven as HAdV-1(8.6%), 30 HAdV-3(37.0%), two HAdV-6(2.5%), and 42 HAdV-7(51.9%). More HAdV-positive samples were collected in August(4.7%, 12/255), September(15.0%, 41/274) and October(6.9%, 17/247), with a peak in September 2018. By combining the results of HAdV phylogenetic analysis with clinical data of patients, there were 77 cases(4.2%, 77/1840;81.1%, 77/95) excluded from nosocomial infections, eight cases representing possible infections transmitted by visitors or attending parents, three cases without sequences that might have been due to infection transmitted by roommates positive for HAdV, one case of a roommate without an HAdV sequence, and six cases that shared highly homologous sequences with those of their roommates, for which nosocomial infections might be considered. In conclusion, genotyping of HAdVs based on phylogenetic analysis combined with clinical information provides a powerful method to distinguish nosocomial infections from community acquired infection, especially when tracing the origins of nosocomial infections.展开更多
To investigate the molecular epidemiology and genetic variation of human adenovirus type 7(HAdV-7)in children with acute respiratory infections(ARI)in China.HAdV-7-positive respiratory samples collected from children ...To investigate the molecular epidemiology and genetic variation of human adenovirus type 7(HAdV-7)in children with acute respiratory infections(ARI)in China.HAdV-7-positive respiratory samples collected from children with ARI in Beijing,Shijiazhuang,Wenzhou and Guangzhou from 2014–2018 were selected for gene amplification and sequence analysis.Fifty-seven HAdV-7 clinical strains with hexon,penton base and fiber gene sequences were obtained.Meanwhile17 strains were selected randomly from different cities for whole genome sequencing.Phylogenetic and variation analyses were performed based on the obtained sequences,HAdV-7 prototype strain Gomen(AY594255),vaccine strains(AY495969 and AY594256)and representative sequences of strains.The phylogenetic trees constructed based on whole genome sequences,major capsid protein genes(hexon,penton base and fiber)and the early genes(E1,E2,E3 and E4)were not completely consistent.The HAdV-7 strains obtained in this study always clustered with most of the circulating strains worldwide from the 1980 s to the present.Compared with the HAdV-7 prototype strain Gomen(AY594255),some amino acid mutations in loop1 and loop2 of hexon and the RGD loop region of the penton base gene were observed.Recombination analysis showed that partial regions of 55 k Da protein and 100 kDa hexon-assembly associated protein genes among all HAdV-7 strains in this study were from HAdV-16 and HAdV-3,respectively.Our study demonstrated the molecular evolution characteristics of HAdV-7 strains circulating in China and provided basic reference data for the prevention,control and vaccine development of HAdV-7.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
Objective Human adenovirus(HAdV)infection is common and can develop to serious conditions with high mortality,yet the mechanism of HAdV infection remains unclear.In the present study,the serum metabolite profiles of H...Objective Human adenovirus(HAdV)infection is common and can develop to serious conditions with high mortality,yet the mechanism of HAdV infection remains unclear.In the present study,the serum metabolite profiles of HAdV-7-infected patients with pneumonia or upper respiratory tract infection(URTI)were explored.Methods In total,35 patients were enrolled in the study following an outbreak of HAdV-7 in the army,of whom 14 had pneumonia and 21 had URTI.Blood samples were collected at the acute stage and at the recovery stage and were analyzed by untargeted metabolomics.Results Over 90% of the differential metabolites identified between the pneumonia patients and URTI patients were lipids and lipid-like molecules,including glycerophospholipids,fatty acyls,and sphingolipids.The metabolic pathways that were significantly enriched were primarily the lipid metabolism pathways,including sphingolipid metabolism,glycerophospholipid metabolism,and linoleic acid metabolism.The sphingolipid metabolism was identified as a significantly differential pathway between the pneumonia patients and URTI patients and between the acute and recovery stages for the pneumonia patients,but not between the acute and recovery stages for the URTI patients.Ceramide and lactosylceramide,involved in sphingolipid metabolism,were significantly higher in the pneumonia patients than in the URTI patients with good discrimination abilities[area under curve(AUC)0.742 and 0.716,respectively;combination AUC 0.801].Conclusion Our results suggested that HAdV modulated lipid metabolism for both the patients with URTI and pneumonia,especially the sphingolipid metabolism involving ceramide and lactosylceramide,which might thus be a potential intervention target in the treatment of HAdV infection.展开更多
Human adenovirus B7(HAdV-B7)causes severe acute lower respiratory tract infections in children.However,neither the child-specific antivirals or vaccines are available,nor the pathogenesis is clear.Autophagy,as part of...Human adenovirus B7(HAdV-B7)causes severe acute lower respiratory tract infections in children.However,neither the child-specific antivirals or vaccines are available,nor the pathogenesis is clear.Autophagy,as part of innate immunity,plays an important role in resistance to viral infection by degrading the virus and promoting the development of innate and adaptive immunity.This study provided evidence that HAdV-B7 infection induced complete autophagic flux,and the pharmacological induction of autophagy decreased HAdV-B7 replication.In this process,the host protein Bcl2-associated athanogene 3(BAG3)mediated autophagy to inhibit the replication of HAdV-B7 by binding to the PPSY structural domain of viral protein pVI through its WW structural domain.These findings further our understanding of the host immune response during viral infection and will help to develop broad anti-HAdV therapies.展开更多
Human adenovirus type 3 （HAdV-3） is widely prevalent all over the world, especially in Asia. The objective of this study is to carry out complete genomic DNA sequencing and the phylogenetic analysis for two strains ...Human adenovirus type 3 （HAdV-3） is widely prevalent all over the world, especially in Asia. The objective of this study is to carry out complete genomic DNA sequencing and the phylogenetic analysis for two strains （Guangzhou01 and Guangzhou02） of HAdV-3 wild virus isolated from South China. Nasopharyngeal secretion aspirate specimens of sick children were inoculated into HEp-2 and HeLa culture tubes, and the cultures were identified by neutralization assay with type-specific reference rabbit antiserum. Type-specific primers were also utilized to confirm the serotype. The restriction fragments of HAdV genome DNA were cloned into pBlueScript SK （ ＋ ） vectors and sequenced, and the 5＇ and 3＇ ends of the linear HAdV-3 genome were directly sequenced with double purified genomic DNA as templates. General features of the HAdV-3 genome sequences were explored by using several bio-software. Phylogenetic analysis was done with MEGA 3.0 software. The genomic sequences of Guangzhou01 and Guangzhou02 possess the same 4 early regions and 5 late regions and have 39 coding sequences and two RNA coding sequences. Other non-coding regions are conservative. Inverted repeats and palindromes were identified in the genome sequences. The genomes of group B human adenovirus as well as HAdV-3 have close phylogenetic relationship with that of chimpanzee adenovirus type 21. The genomic lengths of these two isolated strains are 35 273 bp and 35 269 bp, respectively. The phylogenetic analysis showed that HAdV-B species has some relationship with certain types of chimpanzee adenovirus.展开更多
Introduction:On September 11,2024,a school in the Beijing Economic and Technological Development Area(Jingkai area)reported multiple cases of fever.Public health professionals from the local CDC promptly initiated an ...Introduction:On September 11,2024,a school in the Beijing Economic and Technological Development Area(Jingkai area)reported multiple cases of fever.Public health professionals from the local CDC promptly initiated an on-site epidemiological investigation.Methods:We conducted an initial epidemiological assessment and rapidly identified the index case.Simultaneously,we performed active case finding throughout the school.Throat swab samples were collected from symptomatic individuals and submitted to the laboratory for pathogen testing.Real-time RTPCR was used to screen for a comprehensive panel of respiratory pathogens.We established viral cultures and performed PCR to amplify target gene sequences.Molecular characterization was conducted using gene homology analysis and phylogenetic reconstruction.Results:Active case finding identified 15 fever cases(37.8℃-39.9℃),all from two adjacent classes in the school.Case onset was concentrated between September 9 and 11,with 6 males and 9 females,all aged 6 years.Fourteen throat swab samples were collected,with 10 testing positive for HAdV genes.Six viral strains were successfully isolated through cell culture.Sequence analysis revealed 100% gene homology in the hexon Loop2 region,consistent with HAdV-B3.Furthermore,the penton base gene,hexon gene,and fiber gene of all 6 strains exhibited 100% homology.The penton base gene showed 99.2% homology with the HAdV-B7 reference strain,while the hexon and fiber genes demonstrated 99% and 96.8% homology,respectively,with the HAdV-B3 reference strain.The genotype of all 6 strains was P7H3F3,consistent with the HAdV-B114(P7H3F3)genotype,confirming their identification as HAdVB114.Conclusions:This outbreak was caused by a novel recombinant human adenovirus,HAdV-B114.Given the potential for such emerging HAdV strains to trigger infectious disease outbreaks,enhanced surveillance systems and comprehensive molecular characterization are essential for early detection and effective public health response.展开更多
Introduction:Recent sentinel surveillance has revealed a rising prevalence of human adenovirus type 21(HAdV-21)among HAdV infections in China.This study aimed to elucidate the molecular features of currently circulati...Introduction:Recent sentinel surveillance has revealed a rising prevalence of human adenovirus type 21(HAdV-21)among HAdV infections in China.This study aimed to elucidate the molecular features of currently circulating HAdV-21 strains in China.Methods:Whole-genome sequencing(WGS)was performed on 23 HAdV-21 strains isolated from acute respiratory infection cases,56.5%involving lower respiratory tract infections,across 7 Chinese sentinel surveillance provincial-level administrative divisions(PLADs)(2023-2024).These sequences,along with 50 previously reported HAdV-21 genomes from 6 countries(1956-2019),were integrated into a WGS dataset for comprehensive phylogenetic,genetic variation,and recombination analyses.Results:WGS categorized the HAdV-21 strains into 3 subtypes:HAdV-21a,HAdV-21b,and historical HAdV-21p(isolated in the 1950s).HAdV-21a(1956-2024,involving 5 of the 6 countries)and HAdV-21b(2005-2024,involving 3 of the 6 countries)exhibited extensive spatiotemporal distributions.Recent Chinese strains(2023-2024)belonged to HAdV-21a and HAdV-21b(HAdV-21a/b),showing extremely high genetic homology with Chinese 2019 strains(genetic distance:0.00007)and global strains(distance:<0.00040).Phylogenetic analysis confirmed that HAdV-21a/b shared a common ancestor and maintained a highly conserved genome despite decades of circulation.Sequence variation analysis identified shared and subtype-specific mutations in these two subtypes.Recombination pattern analysis further revealed that HAdV-21a/b acquired an HAdV-3-derived fragment in the E4 region(breakpoint:nt32,843).Conclusions:Recombinant HAdV-21a/b subtypes have co-circulated in China in recent years with remarkable genetic conservation.Enhanced surveillance is essential to quantify associated disease burden and guide targeted prevention and control strategies.展开更多
Background Human adenovirus(HAdV)infection can cause a variety of diseases.It is a major pathogen of pediatric acute respiratory tract infections(ARIs)and can be life-threatening in younger children.We described the e...Background Human adenovirus(HAdV)infection can cause a variety of diseases.It is a major pathogen of pediatric acute respiratory tract infections(ARIs)and can be life-threatening in younger children.We described the epidemiology and subtypes shifting of HAdV among children with ARI in Guangzhou,China.Methods We conducted a retrospective study of 161,079 children diagnosed with acute respiratory illness at the Guangzhou Women and Children’s Medical Center between 2010 and 2021.HAdV specimens were detected by real-time PCR and the hexon gene was used for phylogenetic analysis.Results Before the COVID-19 outbreak in Guangzhou,the annual frequency of adenovirus infection detected during this period ranged from 3.92%to 13.58%,with an epidemic peak every four to fve years.HAdV demonstrated a clear seasonal distribution,with the lowest positivity in March and peaking during summer(July or August)every year.A signifcant increase in HAdV cases was recorded for 2018 and 2019,which coincided with a shift in the dominant HAdV subtype from HAdV-3 to HAdV-7.The latter was associated with a more severe disease compared to HAdV-3.The average mortality proportion for children infected with HAdV from 2016 to 2019 was 0.38%but increased to 20%in severe cases.After COVID-19 emerged,HAdV cases dropped to 2.68%,suggesting that non-pharmaceutical interventions probably reduced the transmission of HAdV in the community.Conclusion Our study provides the foundation for the understanding of the epidemiology of HAdV and its associated risks in children in Southern China.展开更多
Background Outbreaks of severe,acute hepatitis among children have recently attracted global attention.The pathogen causing the outbreak remains unknown,but there is growing evidence that it may be associated with hum...Background Outbreaks of severe,acute hepatitis among children have recently attracted global attention.The pathogen causing the outbreak remains unknown,but there is growing evidence that it may be associated with human adenovirus(HAdV).Data sources A review of adenovirus-related clinical studies,epidemiological studies,etiological studies,and case reports was conducted by reviewers independently.Results HAdV can cause a wide variety of clinical symptoms.In the Mainland of China,HAdV infection accounts for 5.8%–13%of patients with acute respiratory infections,and these infections are mainly caused by species B,C,and E of HAdV.For acute conjunctivitis,39.8%–74.9%of sporadic cases were infected by B and D species of HAdV.Outbreaks of keratoconjunctivitis and pharyngoconjunctival fever related to HAdV infection could be found throughout the country.In pediatric patients with acute gastroenteritis,HAdV-41 was the predominant HAdV type,followed by HAdV species B and C.Several types of HAdV,including HAdV-5,HAdV-7,HAdV-1,and HAdV-2,have previously been reported as potential pathogens associated with HAdV hepatitis in immunocompromised patients.However,few HAdV-related hepatitis cases have been reported in China to date.Conclusions There are no systematic surveillance and clinical studies on HAdV hepatitis in China.Therefore,it is imperative to establish a nationwide HAdV virological surveillance system to collect relevant clinical,epidemiological and virological surveillance data and risk factor information as soon as possible to assess the potential risk of HAdV hepatitis among children.展开更多
obesity and liver steatosis are usually described as related diseases.obesity is regarded as exclusive consequence of an imbalance between food intake and physical exercise,modulated by endocrine and genetic factors.N...obesity and liver steatosis are usually described as related diseases.obesity is regarded as exclusive consequence of an imbalance between food intake and physical exercise,modulated by endocrine and genetic factors.Non-alcoholic fatty liver disease(NAFLD),is a condition whose natural history is related to,but not completely explained by over-nutrition,obesity and insulin resistance.There is evidence that environmental infections,and notably adipogenic adenoviruses(ADV)infections in humans,are associated not only with obesity,which is sufficiently established,but also with allied conditions,such as fatty liver.In order to elucidate the role,if any,of previous ADV36 infection in humans,we investigated association of ADV36-ADV37 seropositivity with obesity and fatty liver in humans.Moreover,the possibility that lifestyle-nutritional intervention in patients with NAFLD and different ADV36 seropositive status,achieves different clinical outcomes on ultrasound bright liver imaging,insulin resistance and obesity was challenged.ADV36 seropositive patientshave a more consistent decrease in insulin resistance,fatty liver severity and body weight in comparison with ADV36 seronegative patients,indicating a greater responsiveness to nutritional intervention.These effects were not dependent on a greater pre-interventional body weight and older age.These results imply that no obvious disadvantage-and,seemingly,that some benefit-is linked to ADV36 seropositivity,at least in NAFLD.ADV36 previous infection can boost weight loss and recovery of insulin sensitivity under interventional treatment.展开更多
Acute respiratory tract infections(ARTIs)are among the leading causes of morbidity and mortality in children worldwide.Human adenovirus(HAdV)infections are estimated to account for at least 5%of pediatric ARTIs.The ci...Acute respiratory tract infections(ARTIs)are among the leading causes of morbidity and mortality in children worldwide.Human adenovirus(HAdV)infections are estimated to account for at least 5%of pediatric ARTIs.The circulated genotypes of HAdV and the correlation between genotype and clinical manifestations in Wuhan,China,before and after the complete relaxation of nonpharmaceutical interventions against severe acute respiratory syndrome coronavirus 2,remain unknown.Here,101 HAdV strains were isolated from throat swab samples collected from hospitalized children with ARTIs who tested positive for HAdV nucleic acid.Of these,sixty-six strains from 2022 to twenty-three strains from 2023 were successfully genotyped and subjected to phylogenetic analysis based on the hexon,penton base,and fiber genes.Six genotypes,B3,C1,C2,C5,C104,and C108 were identified.HAdV-B3(84.85%)was the most prevalent type in 2022,while HAdV-C(86.96%),including C1,C2,C108,and C104,was the most prevalent in 2023.These strains were phylogenetically related to strains from Japan,China,and the United States in recent years.When comparing clinical characteristics,pediatric patients infected with B3,C1,C2,C5,C104,or C108 exhibited similar clinical manifestations,primarily fever and cough,but varying interleukin(IL)-10 levels.In conclusion,from June 2022 to September 2023,the circulated genotypes of HAdV in Wuhan included B3,C1,C2,C108,C5,and C104.The endemic pattern of HAdV in Wuhan,China,shifted from species B as the dominant type in 2022 to species C in 2023.展开更多
Human adenoviruses type 26(HAdV26)and type 35(HAdV35)have increasingly become the choice of adenovirus vectors for vaccine application.However,the population pre-existing immunity to these two adenoviruses in China,wh...Human adenoviruses type 26(HAdV26)and type 35(HAdV35)have increasingly become the choice of adenovirus vectors for vaccine application.However,the population pre-existing immunity to these two adenoviruses in China,which may reduce vaccine efficacy,remains largely unknown.Here,we established micro-neutralizing(MN)assays to investigate the seroprevalence of neutralizing antibodies(nAbs)against HAdV26 and HAdV35 in the general population of Guangdong and Shandong provinces,China.A total of 1184 serum samples were collected,47.0%and 15.8%of which showed HAdV26 and HAdV35 nAb activity,respectively.HAdV26-seropositive individuals tended to have more moderate nAbs titers(201-1000),while HAdV35-seropositive individuals appeared to have more low nAbs titers(72-200).The seropositive rates of HAdV26 and HAdV35 in individuals younger than 20 years old were very low.The seropositive rates of HAdV26 increased with age before 70 years old and decreased thereafter,while HAdV35 seropositive rates did not show similar characteristics.Notably,the seropositive rates and nAb levels of both HAdV26 and HAdV35 were higher in Guangdong Province than in Shandong Province,but did not exert significant differences between males and females.The seroprevalence between HAdV26 and HAdV35 showed little correlation,and no significant cross-neutralizing activity was detected.These results clarified the characteristics of the herd immunity against HAdV26 and HAdV35,and provided information for the rational development and application of HAdV26 and HAdV35 as vaccine vectors in China.展开更多
BACKGROUND： Research of transgene brings hope for gene therapy of various diseases; in addition, some projects have been tested in clinic. Recently, the focus has been to find an ideal vehicle and a suitable therapeu...BACKGROUND： Research of transgene brings hope for gene therapy of various diseases; in addition, some projects have been tested in clinic. Recently, the focus has been to find an ideal vehicle and a suitable therapeutic gene. OBJECTIVE： To explore an effective way to construct recombinant adeno-associated viral vectors expression in human neurnnergen-3 gene. DESIGN： Gene directed cloning. SETTING： Central Laboratory of Northern China Coal Medical College. MATERIALS： DH5a competent bacillus coli strain was provided by Capital Medical University; pCDNA3-NT-3 by professor Chen from Bengbu Medical College; pAAV-Laze, pAAV-Helper, pAAV-RC and pAAV-MCS plasmids by Capital Medical University; HEK293 cells by Cell Center of Basic Medical College of Tongji Medical University. METHODS： NT-3 genes which were selected from pCDNA3-NT-3 plasmids were cloned in pAAV-MCS to form a recombinant adeno-associated viral plasmid （pAAV-NT-3）. pAAV-NT-3, pAAV-RC, pAAV-LacZ and pHelper plasmids were extracted, purified and subjected to enzyme-shearing evaluation. In addition, pAAV-NT-3 and pAAV-LacZ were cotransfected with pHelper and pAAV-RC, respectively into AVV-293 cells with DNA mediated by calcium superphosphate transfection gene; and then, AVV-293 cells were packed into recombinant adeno-associated viral rAAV-NT-3 and rAAV-LacZ. After collection of viral particles, rAAV-LacZ viral stock solution was diluted based on ratio of 10：1 and the mixture was used to infect HT 1080 cells. X-gal stain was used to measure virus titer. MAIN OUTCOME MEASURES： Size of targeted gene fragments, validity of vehicle construction and virus titer. RESULTS： Targeted gene NT-3 was successfully inserted into the relative vehicle pAAV and pAAV-NT-3 was correctly recongnized by enzyme-shearing evaluation. Enzyme-shearing electrophoresis demonstrated that pAAV-NT-3, pAAV-RC, pAAV-LacZ and pHelper plasmids were successfully extracted and purified. β-galactoside staining in situ indicated that LacZ genes were expressed in human fibrosarcoma cells （HT1080） and the recombinant virus titer was measured as 1 ×10^12 virus particles per milliliter. CONCLUSION： Total-length cDNA fragment of NT-3 gene, which is obtained from pCDNA3-NT-3 plasmids, is closely matched to polyclone enzyme-shearing sites of adeno-associated viral vectors, while the combination can be used to construct recombinant adeno-associated viral vectors expression in hNT-3 gene.展开更多
基金supported by the Capital’s Funds for Health Improvement and Research(CFH,shoufa-2022-1G-1131 and shoufa 2022-4G-1133)the High Level Technical Talent Construction Project of Beijing Municipal Health Commission(Discipline Leader-02-20)+1 种基金Beijing Municipal Public Welfare Development and Reform Pilot Project for Medical Research Institutes(JYY2023-10)the Pathogen Spectrum and Host Marker Analysis in Children with Respiratory Tract Infections of Children(Grant 2024-0040).
文摘Objective Recombination events are common and serve as the primary driving force of diverse human adenovirus(HAdV),particularly in children with acute respiratory tract infections(ARIs).Therefore,continual monitoring of these events is essential for effective viral surveillance and control.Methods Respiratory specimens were collected from children with ARIs between January 2022 and December 2023.The penton base,hexon,and fiber genes were amplified from HAdV-positive specimens and sequenced to determine the virus type.In cases with inconsistent typing results,genes were cloned into the pGEM-T vector to detect recombination events.Metagenomic next-generation sequencing(mNGS)was performed to characterize the recombinant HAdV genomes.Results Among 6,771 specimens,277(4.09%,277/6,771)were positvie for HAdV,of which 157(56.68%,157/277)were successfully typed,with HAdV-B3 being the dominant type(91.08%,143/157),and 14(5.05%,14/277)exhibited inconsistent typing results,six of which belonged to species B.The penton base genes of these six specimens were classified as HAdV-B7,whereas their hexon and fiber genes were classified as HAdV-B3,resulting in a recombinant genotype designated P7H3F3,which closely resembled HAdV-B114.Additionally,a partial gene encoding L152/55 kD was identified,which originated from HAdV-B16.Conclusion A novel recombinant,P7H3F3,was identified,containing sequences derived from HAdV-B3 and HAdV-B7,which is similar to HAdV-B114,along with additional sequences from HAdV-B16.
基金supported by National Key Research and Development Program of China(2023YFC2306001)National Natural Science Foundation of China(No.32470141)+1 种基金the Beijing Research Center for Respiratory Infectious Diseases Project(BJRID2025-008)CAMS Innovation Fund for Medical Sciences(CIFMS,NO.2019-I2M-5–026,2022-I2M-CoV19-006).
文摘Human adenovirus type 108(HAdV-108)has been detected in multiple countries,including China,and is associated with severe acute respiratory infection(ARI)in children,with reported fatalities.However,studies on HAdV-108 remain limited.This study aimed to investigate the clinical and genetic characteristics of HAdV-108 in ARI children in China.From 2014 to 2024,6720 respiratory samples were collected from hospitalized children with ARI at ten hospitals across northern and southern China,of which 505(7.51%)tested positive for HAdV.The whole-genome and three major capsid protein genes were amplified and sequenced for bioinformatics analysis,which revealed that among 317 HAdV-isolated samples,21(6.62%)were identified as HAdV-108,ranking third after HAdV-114 and HAdV-7.Clinical analysis of HAdV-108-positive cases showed that the main manifestations were cough and fever.Seven children had gastrointestinal symptoms,and two children without underlying diseases were diagnosed with severe pneumonia.Phylogenetic analysis of wholegenome sequences revealed distinct predominant epidemic branches between domestic and international strains,with one strain obtained in this study forming an independent branch.Hexon protein exhibited the fastest evolution rate,lowest identity,and greatest amino acid variability,while fiber protein displayed the slowest evolution rate,highest identity,and greatest conservation and stability.Compared with the earliest reported HAdV-108 strain,three amino acid deletions were identified in the RGD loop region of penton base protein,resulting in potential structural change.Recombination analysis identified five distinct recombination patterns.In vitro experiments demonstrated that HAdV-108 had proliferation capacity comparable to other species C adenoviruses.In summary,HAdV-108 has persistently circulated in China,causing severe ARIs and concurrent gastrointestinal manifestations.Cluster3 was the predominant epidemic branch in China.HAdV-108 exhibited significant intratype genetic variation,with random and diverse recombination events.
基金supported by the Guangzhou Science and Technology Program-Zhongnanshan Medical Foundation of Guangdong Province(202102010359-ZNSA-2020003)the Emergency Key Program of Guangzhou Laboratory(EKPG21-13)+3 种基金the National Natural Science Foundation of China(81970003,31900877)the Natural Science Foundation of Guangdong Province of China(2018A030310401)Guangdong-Hong Kong-Macao Joint Laboratory of Respiratory Infectious Disease(GHMJLRID-Z-202109)the Special Project for COVID-19 Prevention and Control of Zhongnanshan Medical Foundation of Guangdong Province(ZNSA-2020012)。
文摘During 2018–2019,a severe human adenovirus(HAdV)infection outbreak occurred in southern China.Here,we screened 18 respiratory pathogens in 1704 children(≤14 years old)hospitalized with acute respiratory illness in Guangzhou,China,in 2019.In total,151 patients had positive HAdV test results;34.4%(52/151)of them exhibited severe illness.HAdV infection occurred throughout the year,with a peak in summer.The median patient age was 3.0(interquartile range:1.1–5.0)years.Patients with severe HAdV infection exhibited increases in12 clinical indexes(P≤0.019)and decreases in four indexes(P≤0.007),compared with patients exhibiting nonsevere infection.No significant differences were found in age or sex distribution according to HAdV infection severity(P>0.05);however,the distributions of comorbid disease and HAdV co-infection differed according to HAdV infection severity(P<0.05).The main epidemic types were HAdV-3(47.0%,71/151)and HAdV-7(46.4%,70/151).However,the severe illness rate was significantly higher in patients with HAdV-7(51.4%)than in patients with HAdV-3(19.7%)and other types of HAdV(20%)(P<0.001).Sequencing analysis of genomes/capsid genes of 13 HAdV-7 isolates revealed high similarity to previous Chinese isolates.A representative HAdV-7isolate exhibited a similar proliferation curve to the curve described for the epidemic HAdV-3 strain Guangzhou01(accession no.DQ099432)(P>0.05);the HAdV-7 isolate exhibited stronger virulence and infectivity,compared with HAdV-3(P<0.001).Overall,comorbid disease,HAdV co-infection,and high virulence and infectivity of HAdV-7 were critical risk factors for severe HAdV infection;these data can facilitate treatment,control,and prevention of HAdV infection.
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 the National Key Research and Development Program of China (2018YFC1200100)the Guangzhou Science and Technology Program Key Project, China (201803040004)+2 种基金the National Science and Technology Major Project of China (2017ZX10103011003, 2018ZX10102001)the National Natural Science Foundation of China, China (31570163)the Youth Project of State Key Laboratory of Respiratory Disease, China (SKLRD-QN-201713)。
文摘Human adenoviruses(HAdVs) commonly cause many diseases such as respiratory diseases, gastroenteritis, cystitis worldwide. HAdV-3,-7,-4 and emergent HAdV-55 and HAdV-14 are the most important types causing severe respiratory diseases. There is no effective drug available for clinical treatment, and no vaccine available for the general population.Therefore, it is important to investigate the seroprevalence against HAdV for developing novel vaccines and vectors. In this study, we investigated the seroprevalence and titer levels of neutralizing antibodies(NAb) against HAdV-3,-4,-7,-14,-55,and-11 in total 278 healthy populations between 0 months and 49 years of age(228 children and 50 adults) from Guangzhou. In children under the age of 18 years, the seropositive rates were significantly increased against HAdV-3 at12.07%, 33.96%, and 64.29% and against HAdV-7 at 0%, 18.87%, and 19.05% in age groups of 1–2, 3–5, and 6–17 years,respectively. The seroprevalence was very low(0% - 8.1%) for all other four types. In adults aged between 18 and49 years, HAdV-3,-4, and-7(> 50.00%) were the most common types, followed by HAdV-14(38.00%),-55(34.00%),and-11(24.00%). Adults tended to have high NAb titers against HAdV-4 and-55. HAdV-55-seropositive donors tended to be HAdV-11-and HAdV-14-seropositive. These results indicated the low level of herd immunity against all six HAdV types in young children, and HAdV-14,-55,-11 in adults from Guangzhou City. Our findings demonstrate the importance of monitoring HAdV types and developing vaccines against HAdV for children and adults.
基金This work was supported by grants from the National Key Research and Development Program of China(2018YFE0204503)Natural Science Foundation of Guangdong Province(2021A1515010788 and 2018B030312010)the Guangzhou Healthcare Collaborative Innovation Major Project(201803040004 and 201803040007)。
文摘Human adenovirus type 55(HAdV-B55) is a re-emergent acute respiratory disease pathogen that causes adult communityacquired pneumonia(CAP). Previous studies have shown that the receptor of HAdV-B14, which genome is highly similar with HAdV-B55, is human Desmoglein 2(DSG2). However, whether the receptor of HAdV-B55 is DSG2 is undetermined because there are three amino acid mutations in the fiber gene between HAdV-B14 and HAdV-B55. Here, firstly we found the 3T3 cells, a mouse embryo fibroblast rodent cell line which does not express human DSG2, were able to be infected by HAdV-B55 after transfected with pcDNA3.1-DSG2, while normal 3T3 cells were still unsusceptible to HAdV-B55 infection. Next, A549 cells with h DSG2 knock-down by siRNA were hard to be infected by HAdV-B3/-B14/-B55, while the control siRNA group was still able to be infected by all these types of HAdVs. Finally, immunofluorescence confocal microscopy indicated visually that Cy3-conjugated HAdV-B55 viruses entered A549 cells by binding to DSG2 protein.Therefore, DSG2 is a major receptor of HAdV-B55 causing adult CAP. Our finding is important for better understanding of interactions between adenoviruses and host cells and may shed light on the development of new drugs that can interfere with these processes as well as for the development of potent prophylactic vaccines.
基金supported by grants from Beijing Municipal Commission of Health and Family (No. 2060399 PXM2017_026268_00005_ 00254486)The Pediatric Medical Coordinated Development Center of the Beijing Hospitals Authority (No. XTZD20180505)。
文摘Nosocomial infections are common in pediatric patients and can be fatal in infants and immunocompromised patients. In September 2018, a high positive rate of human adenovirus HAdV was occurred among hospitalized children in the Children's Hospital Affiliated to the Capital Institute of Paediatrics in Beijing. To investigate whether this outbreak of HAdV was related to nosocomial infections or the result of community infections, we collected respiratory specimens from patients with acute respiratory infections in a respiratory ward during June to December 2018, and screened for respiratory viruses. Among 1,840 cases included, 95(5.2%, 95/1840) were positive for HAdV and 81 were genotyped based on phylogenetic analysis, including seven as HAdV-1(8.6%), 30 HAdV-3(37.0%), two HAdV-6(2.5%), and 42 HAdV-7(51.9%). More HAdV-positive samples were collected in August(4.7%, 12/255), September(15.0%, 41/274) and October(6.9%, 17/247), with a peak in September 2018. By combining the results of HAdV phylogenetic analysis with clinical data of patients, there were 77 cases(4.2%, 77/1840;81.1%, 77/95) excluded from nosocomial infections, eight cases representing possible infections transmitted by visitors or attending parents, three cases without sequences that might have been due to infection transmitted by roommates positive for HAdV, one case of a roommate without an HAdV sequence, and six cases that shared highly homologous sequences with those of their roommates, for which nosocomial infections might be considered. In conclusion, genotyping of HAdVs based on phylogenetic analysis combined with clinical information provides a powerful method to distinguish nosocomial infections from community acquired infection, especially when tracing the origins of nosocomial infections.
基金funded by the Key Technology R&D Program of China(grant numbers2017ZX10103004-004,2017ZX10104001-005-010)National Natural Science Foundation of China(grant number 82072266)CAMS Innovation Fund for Medical Sciences(CIFMS),(grant number 2019-I2M-5-026)。
文摘To investigate the molecular epidemiology and genetic variation of human adenovirus type 7(HAdV-7)in children with acute respiratory infections(ARI)in China.HAdV-7-positive respiratory samples collected from children with ARI in Beijing,Shijiazhuang,Wenzhou and Guangzhou from 2014–2018 were selected for gene amplification and sequence analysis.Fifty-seven HAdV-7 clinical strains with hexon,penton base and fiber gene sequences were obtained.Meanwhile17 strains were selected randomly from different cities for whole genome sequencing.Phylogenetic and variation analyses were performed based on the obtained sequences,HAdV-7 prototype strain Gomen(AY594255),vaccine strains(AY495969 and AY594256)and representative sequences of strains.The phylogenetic trees constructed based on whole genome sequences,major capsid protein genes(hexon,penton base and fiber)and the early genes(E1,E2,E3 and E4)were not completely consistent.The HAdV-7 strains obtained in this study always clustered with most of the circulating strains worldwide from the 1980 s to the present.Compared with the HAdV-7 prototype strain Gomen(AY594255),some amino acid mutations in loop1 and loop2 of hexon and the RGD loop region of the penton base gene were observed.Recombination analysis showed that partial regions of 55 k Da protein and 100 kDa hexon-assembly associated protein genes among all HAdV-7 strains in this study were from HAdV-16 and HAdV-3,respectively.Our study demonstrated the molecular evolution characteristics of HAdV-7 strains circulating in China and provided basic reference data for the prevention,control and vaccine development of HAdV-7.
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(No.82073617)Joint Research Fund for Beijing Natural Science Foundation and Haidian Original Innovation(No.L202007)+1 种基金Fundamental Research Funds for the Central Universities and Peking University Health Science Center(No.BMU2021YJ041)Peking University Medicine Fund of Fostering Young Scholars'Scientific&Technological Innovation(No.BMU2021PY005).
文摘Objective Human adenovirus(HAdV)infection is common and can develop to serious conditions with high mortality,yet the mechanism of HAdV infection remains unclear.In the present study,the serum metabolite profiles of HAdV-7-infected patients with pneumonia or upper respiratory tract infection(URTI)were explored.Methods In total,35 patients were enrolled in the study following an outbreak of HAdV-7 in the army,of whom 14 had pneumonia and 21 had URTI.Blood samples were collected at the acute stage and at the recovery stage and were analyzed by untargeted metabolomics.Results Over 90% of the differential metabolites identified between the pneumonia patients and URTI patients were lipids and lipid-like molecules,including glycerophospholipids,fatty acyls,and sphingolipids.The metabolic pathways that were significantly enriched were primarily the lipid metabolism pathways,including sphingolipid metabolism,glycerophospholipid metabolism,and linoleic acid metabolism.The sphingolipid metabolism was identified as a significantly differential pathway between the pneumonia patients and URTI patients and between the acute and recovery stages for the pneumonia patients,but not between the acute and recovery stages for the URTI patients.Ceramide and lactosylceramide,involved in sphingolipid metabolism,were significantly higher in the pneumonia patients than in the URTI patients with good discrimination abilities[area under curve(AUC)0.742 and 0.716,respectively;combination AUC 0.801].Conclusion Our results suggested that HAdV modulated lipid metabolism for both the patients with URTI and pneumonia,especially the sphingolipid metabolism involving ceramide and lactosylceramide,which might thus be a potential intervention target in the treatment of HAdV infection.
基金This work was supported by the National Natural Science Foundation of China(No.82072266)the CAMS Innovation Fund for Medical Sciences(No.2019-I2M-5-026).
文摘Human adenovirus B7(HAdV-B7)causes severe acute lower respiratory tract infections in children.However,neither the child-specific antivirals or vaccines are available,nor the pathogenesis is clear.Autophagy,as part of innate immunity,plays an important role in resistance to viral infection by degrading the virus and promoting the development of innate and adaptive immunity.This study provided evidence that HAdV-B7 infection induced complete autophagic flux,and the pharmacological induction of autophagy decreased HAdV-B7 replication.In this process,the host protein Bcl2-associated athanogene 3(BAG3)mediated autophagy to inhibit the replication of HAdV-B7 by binding to the PPSY structural domain of viral protein pVI through its WW structural domain.These findings further our understanding of the host immune response during viral infection and will help to develop broad anti-HAdV therapies.
文摘Human adenovirus type 3 （HAdV-3） is widely prevalent all over the world, especially in Asia. The objective of this study is to carry out complete genomic DNA sequencing and the phylogenetic analysis for two strains （Guangzhou01 and Guangzhou02） of HAdV-3 wild virus isolated from South China. Nasopharyngeal secretion aspirate specimens of sick children were inoculated into HEp-2 and HeLa culture tubes, and the cultures were identified by neutralization assay with type-specific reference rabbit antiserum. Type-specific primers were also utilized to confirm the serotype. The restriction fragments of HAdV genome DNA were cloned into pBlueScript SK （ ＋ ） vectors and sequenced, and the 5＇ and 3＇ ends of the linear HAdV-3 genome were directly sequenced with double purified genomic DNA as templates. General features of the HAdV-3 genome sequences were explored by using several bio-software. Phylogenetic analysis was done with MEGA 3.0 software. The genomic sequences of Guangzhou01 and Guangzhou02 possess the same 4 early regions and 5 late regions and have 39 coding sequences and two RNA coding sequences. Other non-coding regions are conservative. Inverted repeats and palindromes were identified in the genome sequences. The genomes of group B human adenovirus as well as HAdV-3 have close phylogenetic relationship with that of chimpanzee adenovirus type 21. The genomic lengths of these two isolated strains are 35 273 bp and 35 269 bp, respectively. The phylogenetic analysis showed that HAdV-B species has some relationship with certain types of chimpanzee adenovirus.
文摘Introduction:On September 11,2024,a school in the Beijing Economic and Technological Development Area(Jingkai area)reported multiple cases of fever.Public health professionals from the local CDC promptly initiated an on-site epidemiological investigation.Methods:We conducted an initial epidemiological assessment and rapidly identified the index case.Simultaneously,we performed active case finding throughout the school.Throat swab samples were collected from symptomatic individuals and submitted to the laboratory for pathogen testing.Real-time RTPCR was used to screen for a comprehensive panel of respiratory pathogens.We established viral cultures and performed PCR to amplify target gene sequences.Molecular characterization was conducted using gene homology analysis and phylogenetic reconstruction.Results:Active case finding identified 15 fever cases(37.8℃-39.9℃),all from two adjacent classes in the school.Case onset was concentrated between September 9 and 11,with 6 males and 9 females,all aged 6 years.Fourteen throat swab samples were collected,with 10 testing positive for HAdV genes.Six viral strains were successfully isolated through cell culture.Sequence analysis revealed 100% gene homology in the hexon Loop2 region,consistent with HAdV-B3.Furthermore,the penton base gene,hexon gene,and fiber gene of all 6 strains exhibited 100% homology.The penton base gene showed 99.2% homology with the HAdV-B7 reference strain,while the hexon and fiber genes demonstrated 99% and 96.8% homology,respectively,with the HAdV-B3 reference strain.The genotype of all 6 strains was P7H3F3,consistent with the HAdV-B114(P7H3F3)genotype,confirming their identification as HAdVB114.Conclusions:This outbreak was caused by a novel recombinant human adenovirus,HAdV-B114.Given the potential for such emerging HAdV strains to trigger infectious disease outbreaks,enhanced surveillance systems and comprehensive molecular characterization are essential for early detection and effective public health response.
基金Supported by the National Key R&D Program of China(2022YFC2305303)and the Health and Wellness Sector Research Program of Gansu(GSWSON2021-010).
文摘Introduction:Recent sentinel surveillance has revealed a rising prevalence of human adenovirus type 21(HAdV-21)among HAdV infections in China.This study aimed to elucidate the molecular features of currently circulating HAdV-21 strains in China.Methods:Whole-genome sequencing(WGS)was performed on 23 HAdV-21 strains isolated from acute respiratory infection cases,56.5%involving lower respiratory tract infections,across 7 Chinese sentinel surveillance provincial-level administrative divisions(PLADs)(2023-2024).These sequences,along with 50 previously reported HAdV-21 genomes from 6 countries(1956-2019),were integrated into a WGS dataset for comprehensive phylogenetic,genetic variation,and recombination analyses.Results:WGS categorized the HAdV-21 strains into 3 subtypes:HAdV-21a,HAdV-21b,and historical HAdV-21p(isolated in the 1950s).HAdV-21a(1956-2024,involving 5 of the 6 countries)and HAdV-21b(2005-2024,involving 3 of the 6 countries)exhibited extensive spatiotemporal distributions.Recent Chinese strains(2023-2024)belonged to HAdV-21a and HAdV-21b(HAdV-21a/b),showing extremely high genetic homology with Chinese 2019 strains(genetic distance:0.00007)and global strains(distance:<0.00040).Phylogenetic analysis confirmed that HAdV-21a/b shared a common ancestor and maintained a highly conserved genome despite decades of circulation.Sequence variation analysis identified shared and subtype-specific mutations in these two subtypes.Recombination pattern analysis further revealed that HAdV-21a/b acquired an HAdV-3-derived fragment in the E4 region(breakpoint:nt32,843).Conclusions:Recombinant HAdV-21a/b subtypes have co-circulated in China in recent years with remarkable genetic conservation.Enhanced surveillance is essential to quantify associated disease burden and guide targeted prevention and control strategies.
基金supported by the National Natural Science Foundation of China:[grant number 82072264]Natural Science Foundation of Guangdong Province,China:[grant number 2021A1515011071]+2 种基金Guangzhou School(Institute)Joint Funding Project(No.202102010202)Municipal Science and Technology Bureau Foundation of Guangzhou(201803040004)Guangzhou Basic Research Program Co-funded by Zhongnanshan Medical Foundation of Guangdong Province:[grant number 202102010364,ZNSA-2020003].
文摘Background Human adenovirus(HAdV)infection can cause a variety of diseases.It is a major pathogen of pediatric acute respiratory tract infections(ARIs)and can be life-threatening in younger children.We described the epidemiology and subtypes shifting of HAdV among children with ARI in Guangzhou,China.Methods We conducted a retrospective study of 161,079 children diagnosed with acute respiratory illness at the Guangzhou Women and Children’s Medical Center between 2010 and 2021.HAdV specimens were detected by real-time PCR and the hexon gene was used for phylogenetic analysis.Results Before the COVID-19 outbreak in Guangzhou,the annual frequency of adenovirus infection detected during this period ranged from 3.92%to 13.58%,with an epidemic peak every four to fve years.HAdV demonstrated a clear seasonal distribution,with the lowest positivity in March and peaking during summer(July or August)every year.A signifcant increase in HAdV cases was recorded for 2018 and 2019,which coincided with a shift in the dominant HAdV subtype from HAdV-3 to HAdV-7.The latter was associated with a more severe disease compared to HAdV-3.The average mortality proportion for children infected with HAdV from 2016 to 2019 was 0.38%but increased to 20%in severe cases.After COVID-19 emerged,HAdV cases dropped to 2.68%,suggesting that non-pharmaceutical interventions probably reduced the transmission of HAdV in the community.Conclusion Our study provides the foundation for the understanding of the epidemiology of HAdV and its associated risks in children in Southern China.
基金Key Public Health Projects of the National Health Commission(ZDGW21-131031103000180005)The Key Technologies R&D Program of the Ministry of Science and Technology(2018ZX10713002).
文摘Background Outbreaks of severe,acute hepatitis among children have recently attracted global attention.The pathogen causing the outbreak remains unknown,but there is growing evidence that it may be associated with human adenovirus(HAdV).Data sources A review of adenovirus-related clinical studies,epidemiological studies,etiological studies,and case reports was conducted by reviewers independently.Results HAdV can cause a wide variety of clinical symptoms.In the Mainland of China,HAdV infection accounts for 5.8%–13%of patients with acute respiratory infections,and these infections are mainly caused by species B,C,and E of HAdV.For acute conjunctivitis,39.8%–74.9%of sporadic cases were infected by B and D species of HAdV.Outbreaks of keratoconjunctivitis and pharyngoconjunctival fever related to HAdV infection could be found throughout the country.In pediatric patients with acute gastroenteritis,HAdV-41 was the predominant HAdV type,followed by HAdV species B and C.Several types of HAdV,including HAdV-5,HAdV-7,HAdV-1,and HAdV-2,have previously been reported as potential pathogens associated with HAdV hepatitis in immunocompromised patients.However,few HAdV-related hepatitis cases have been reported in China to date.Conclusions There are no systematic surveillance and clinical studies on HAdV hepatitis in China.Therefore,it is imperative to establish a nationwide HAdV virological surveillance system to collect relevant clinical,epidemiological and virological surveillance data and risk factor information as soon as possible to assess the potential risk of HAdV hepatitis among children.
文摘obesity and liver steatosis are usually described as related diseases.obesity is regarded as exclusive consequence of an imbalance between food intake and physical exercise,modulated by endocrine and genetic factors.Non-alcoholic fatty liver disease(NAFLD),is a condition whose natural history is related to,but not completely explained by over-nutrition,obesity and insulin resistance.There is evidence that environmental infections,and notably adipogenic adenoviruses(ADV)infections in humans,are associated not only with obesity,which is sufficiently established,but also with allied conditions,such as fatty liver.In order to elucidate the role,if any,of previous ADV36 infection in humans,we investigated association of ADV36-ADV37 seropositivity with obesity and fatty liver in humans.Moreover,the possibility that lifestyle-nutritional intervention in patients with NAFLD and different ADV36 seropositive status,achieves different clinical outcomes on ultrasound bright liver imaging,insulin resistance and obesity was challenged.ADV36 seropositive patientshave a more consistent decrease in insulin resistance,fatty liver severity and body weight in comparison with ADV36 seronegative patients,indicating a greater responsiveness to nutritional intervention.These effects were not dependent on a greater pre-interventional body weight and older age.These results imply that no obvious disadvantage-and,seemingly,that some benefit-is linked to ADV36 seropositivity,at least in NAFLD.ADV36 previous infection can boost weight loss and recovery of insulin sensitivity under interventional treatment.
基金supported by the projects of National Virus Resource Center(NVRC-PY-03,E1YZ020501)Natural Science Foundation of Hubei Province(2022CFB564)+4 种基金Foundation of Hubei Provincial Health Commission(WJ2023M108)National Basic Science Data Sharing Platform(no.2018ZX10101004)National Basic Science Data Sharing Service Platform(no.NBSDC-DB-13)the International Cooperation Base of Hubei Province for Infection and Immunitysupported by Outstanding Medical Young Scholars of Hubei Province and Wuhan Young and Middle-aged Medical Backbone Talent Program.
文摘Acute respiratory tract infections(ARTIs)are among the leading causes of morbidity and mortality in children worldwide.Human adenovirus(HAdV)infections are estimated to account for at least 5%of pediatric ARTIs.The circulated genotypes of HAdV and the correlation between genotype and clinical manifestations in Wuhan,China,before and after the complete relaxation of nonpharmaceutical interventions against severe acute respiratory syndrome coronavirus 2,remain unknown.Here,101 HAdV strains were isolated from throat swab samples collected from hospitalized children with ARTIs who tested positive for HAdV nucleic acid.Of these,sixty-six strains from 2022 to twenty-three strains from 2023 were successfully genotyped and subjected to phylogenetic analysis based on the hexon,penton base,and fiber genes.Six genotypes,B3,C1,C2,C5,C104,and C108 were identified.HAdV-B3(84.85%)was the most prevalent type in 2022,while HAdV-C(86.96%),including C1,C2,C108,and C104,was the most prevalent in 2023.These strains were phylogenetically related to strains from Japan,China,and the United States in recent years.When comparing clinical characteristics,pediatric patients infected with B3,C1,C2,C5,C104,or C108 exhibited similar clinical manifestations,primarily fever and cough,but varying interleukin(IL)-10 levels.In conclusion,from June 2022 to September 2023,the circulated genotypes of HAdV in Wuhan included B3,C1,C2,C108,C5,and C104.The endemic pattern of HAdV in Wuhan,China,shifted from species B as the dominant type in 2022 to species C in 2023.
基金supported by the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB29050701)the Emergency Key Program of Guangzhou (EKPG21-20)+2 种基金the China Evergrande Group funding for SARS-Co V-2 (2020GIRHHMS22)the Zhongnanshan Medical Foundation of Guangdong Province (ZNSA-2022009)the China Postdoctoral Science Foundation (2020M682942)
文摘Human adenoviruses type 26(HAdV26)and type 35(HAdV35)have increasingly become the choice of adenovirus vectors for vaccine application.However,the population pre-existing immunity to these two adenoviruses in China,which may reduce vaccine efficacy,remains largely unknown.Here,we established micro-neutralizing(MN)assays to investigate the seroprevalence of neutralizing antibodies(nAbs)against HAdV26 and HAdV35 in the general population of Guangdong and Shandong provinces,China.A total of 1184 serum samples were collected,47.0%and 15.8%of which showed HAdV26 and HAdV35 nAb activity,respectively.HAdV26-seropositive individuals tended to have more moderate nAbs titers(201-1000),while HAdV35-seropositive individuals appeared to have more low nAbs titers(72-200).The seropositive rates of HAdV26 and HAdV35 in individuals younger than 20 years old were very low.The seropositive rates of HAdV26 increased with age before 70 years old and decreased thereafter,while HAdV35 seropositive rates did not show similar characteristics.Notably,the seropositive rates and nAb levels of both HAdV26 and HAdV35 were higher in Guangdong Province than in Shandong Province,but did not exert significant differences between males and females.The seroprevalence between HAdV26 and HAdV35 showed little correlation,and no significant cross-neutralizing activity was detected.These results clarified the characteristics of the herd immunity against HAdV26 and HAdV35,and provided information for the rational development and application of HAdV26 and HAdV35 as vaccine vectors in China.
文摘BACKGROUND： Research of transgene brings hope for gene therapy of various diseases; in addition, some projects have been tested in clinic. Recently, the focus has been to find an ideal vehicle and a suitable therapeutic gene. OBJECTIVE： To explore an effective way to construct recombinant adeno-associated viral vectors expression in human neurnnergen-3 gene. DESIGN： Gene directed cloning. SETTING： Central Laboratory of Northern China Coal Medical College. MATERIALS： DH5a competent bacillus coli strain was provided by Capital Medical University; pCDNA3-NT-3 by professor Chen from Bengbu Medical College; pAAV-Laze, pAAV-Helper, pAAV-RC and pAAV-MCS plasmids by Capital Medical University; HEK293 cells by Cell Center of Basic Medical College of Tongji Medical University. METHODS： NT-3 genes which were selected from pCDNA3-NT-3 plasmids were cloned in pAAV-MCS to form a recombinant adeno-associated viral plasmid （pAAV-NT-3）. pAAV-NT-3, pAAV-RC, pAAV-LacZ and pHelper plasmids were extracted, purified and subjected to enzyme-shearing evaluation. In addition, pAAV-NT-3 and pAAV-LacZ were cotransfected with pHelper and pAAV-RC, respectively into AVV-293 cells with DNA mediated by calcium superphosphate transfection gene; and then, AVV-293 cells were packed into recombinant adeno-associated viral rAAV-NT-3 and rAAV-LacZ. After collection of viral particles, rAAV-LacZ viral stock solution was diluted based on ratio of 10：1 and the mixture was used to infect HT 1080 cells. X-gal stain was used to measure virus titer. MAIN OUTCOME MEASURES： Size of targeted gene fragments, validity of vehicle construction and virus titer. RESULTS： Targeted gene NT-3 was successfully inserted into the relative vehicle pAAV and pAAV-NT-3 was correctly recongnized by enzyme-shearing evaluation. Enzyme-shearing electrophoresis demonstrated that pAAV-NT-3, pAAV-RC, pAAV-LacZ and pHelper plasmids were successfully extracted and purified. β-galactoside staining in situ indicated that LacZ genes were expressed in human fibrosarcoma cells （HT1080） and the recombinant virus titer was measured as 1 ×10^12 virus particles per milliliter. CONCLUSION： Total-length cDNA fragment of NT-3 gene, which is obtained from pCDNA3-NT-3 plasmids, is closely matched to polyclone enzyme-shearing sites of adeno-associated viral vectors, while the combination can be used to construct recombinant adeno-associated viral vectors expression in hNT-3 gene.
