scholarly journals Enhancement of Enteric Adenovirus Cultivation by Viral Transactivator Proteins

2010 ◽  
Vol 76 (8) ◽  
pp. 2509-2516 ◽  
Author(s):  
Misoon Kim ◽  
Mi Young Lim ◽  
GwangPyo Ko
Keyword(s):  
Cell Line ◽  
Cytopathic Effect ◽  
Cell Cultivation ◽  
Promoter Regions ◽  
Viral Dna ◽  
Viral Transactivator ◽  
Food Borne ◽  
B Virus ◽  
Enteric Adenovirus ◽  
Enteric Adenoviruses

ABSTRACT Human enteric adenoviruses (HAdVs; serotypes 40 and 41) are important waterborne and food-borne pathogens. However, HAdVs are fastidious, are difficult to cultivate, and do not produce a clear cytopathic effect during cell culture within a reasonable time. Thus, we examined whether the viral transactivator proteins cytomegalovirus (CMV) IE1 and hepatitis B virus (HBV) X promoted the multiplication of HAdVs. Additionally, we constructed a new 293 cell line expressing CMV IE1 protein for cultivation assays. We analyzed the nucleic acid sequences of the promoter regions of both E1A and hexon genes, which are considered to be the most important regions for HAdV replication. Expression of either HBV X or CMV IE1 protein significantly increased the promoter activities of E1A and hexon genes of HAdVs by as much as 14-fold during cell cultivation. The promotion of HAdV expression was confirmed by increased levels of both adenoviral DNA and mRNA expression. Finally, the newly developed 293 cell line expressing CMV IE1 protein showed an increase in viral DNA ranging from 574% to 619% compared with the conventional 293 cell line. These results suggest that the newly constructed cell line could be useful for efficient cultivation and research of fastidious HAdVs.

scholarly journals Improved growth of enteric adenovirus type 40 in a modified cell line that can no longer respond to interferon stimulation

2007 ◽  
Vol 88 (1) ◽  
pp. 71-76 ◽  
Author(s):  
Victoria Sherwood ◽  
Hans-Gerhard Burgert ◽  
Yun-Hsiang Chen ◽  
Sandeep Sanghera ◽  
Socrates Katafigiotis ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Adenovirus Type ◽  
Simian Virus ◽  
Response Analysis ◽  
V Protein ◽  
Simian Virus 5 ◽  
293 Cells ◽  
Enteric Adenovirus ◽  
Enteric Adenoviruses

Human enteric adenoviruses propagate poorly in conventional human cell lines used to grow other adenovirus serotypes. As human enteric adenoviruses have a defect in counteracting the cellular interferon (IFN) response in cell culture, to aid in growth of the virus, a 293-based cell line defective in its ability to respond to IFN was constructed. This cell line (293-SV5/V) constitutively expresses V-protein of the paramyxovirus Simian virus 5, which degrades the signal transducer and activator of transcription 1 (STAT1) and thereby prevents the STAT1-mediated IFN response. Analysis of human enteric adenovirus type 40 (HAdV-40)-infected 293-SV5/V cells compared with parental 293 cells shows that the recombinant line allows more rapid production of virus and results in higher titres. These results suggest that the defect in HAdV-40 in counteracting the IFN response can be overcome at least partially through the use of 293-SV5/V cell lines.

Lower hepatitis B virus productivity in HBeAg negative patients is not due to accumulation of mutations within viral DNA regulatory regions

2009 ◽  
Vol 47 (01) ◽  
Author(s):  
A Köpke ◽  
T Volz ◽  
M Lütgehetmann ◽  
AW Lohse ◽  
J Petersen ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Viral Dna ◽  
Regulatory Regions ◽  
B Virus

Detection of enteric adenoviruses in south African waters using gene probes

1995 ◽  
Vol 31 (5-6) ◽  
pp. 345-350 ◽  
Author(s):  
B. Genthe ◽  
M. Gericke ◽  
B. Bateman ◽  
N. Mjoli ◽  
R. Kfir
Keyword(s):  
South African ◽  
Water Samples ◽  
Cellulose Fibre ◽  
Gene Probe ◽  
Viral Dna ◽  
Virus Particles ◽  
Gene Probes ◽  
Probe Hybridization ◽  
Enteric Adenoviruses ◽  
Effect Of Chlorine

Gene probes developed locally for both enteric Adenoviruses 40 and 41 were used to determine whether these viruses were present in both raw and treated waters. Approximately sixty water samples were concentrated by ultrafiltration and analysed directly for the presence of enteric adenoviruses. Three pretreatment techniques, namely sephadex columns, cellulose fibre and GenecleanTM were tested for the removal of inhibitory substances from concentrated water samples. The effect of chlorine treatment on viral detection using gene probe hybridization was also examined by exposing adenoviruses to chlorine concentrations of up to 20mg/l for 1 hour. Enteric adenoviruses were detected in up to 59% of both raw and treated waters analysed. Cellulose fibre and GenecleanTM were found to successfully remove inhibitory substances from concentrated raw waters. Viral DNA was detected after exposure to a range of chlorine concentrations indicating that the viruses detected in the treated waters may have been inactivated virus particles.

Detection and molecular characterization of enteric adenovirus in treated wastewater in the Brazilian Federal District

2021 ◽  
Vol 3 (7) ◽  
Author(s):  
T. S. C. Quintão ◽  
F. G. Silva ◽  
A. L. Pereira ◽  
W. N. Araújo ◽  
P. M. Oliveira ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Wastewater Treatment ◽  
Wastewater Treatment Plants ◽  
Federal District ◽  
Treated Wastewater ◽  
Conventional Pcr ◽  
Rainfall Index ◽  
Enteric Adenovirus ◽  
Enteric Adenoviruses

AbstractHuman enteric viruses, such as enteric adenoviruses (HAdV), are known to be involved with gastrointestinal disorders, especially acute gastroenteritis. Several studies have used HAdV as an indicator of water quality, since they are considered highly stable and widely distributed viruses in water matrices. The aim of this study was to detect and genotype HAdVs in water matrices impacted by discharges of treated effluents from wastewater treatment plants (WWTPs). Wastewater treatment plants from the sanitary system of the Brazilian Federal District were assessed in 2018 and 2019. Samples were collected upstream and downstream from discharge points for each WWTP. Viral concentration based on adsorption-elution and conventional PCR was used for molecular detection, and positive samples were sequenced for phylogenetic analysis. Pluviosity data for the period in which the samples were collected were obtained. Our results demonstrated the presence of HAdVs in 27.2% (61/224) of the samples. The positivity was significantly higher in downstream samples compared to upstream. Moreover, the HAdV positivity was higher in downstream samples collected from receiving water bodies impacted by secondary-level WWTPs in comparison with those impacted by tertiary-level WWTPs. Phylogenetic analysis demonstrated the presence of genotypes 40 and 41, with prevalence of HAdV genotype 41. Despite the predominance of HAdV-41, an increasing frequency of the HAdV-40 was associated with higher pluviosity. In conclusion, this study is the first documentation in the Brazilian Federal District dealing with the prevalence and diversity of HAdVs in several WWTP, along with their correlation with rainfall index.

scholarly journals Creation of a Six-fingered Artificial Transcription Factor That Represses the Hepatitis B Virus HBx Gene Integrated into a Human Hepatocellular Carcinoma Cell Line

2012 ◽  
Vol 18 (4) ◽  
pp. 378-387 ◽  
Author(s):  
Xinghui Zhao ◽  
Zhanzhong Zhao ◽  
Junwei Guo ◽  
Peitang Huang ◽  
Xudong Zhu ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Transcription Factor ◽  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Cell Line ◽  
Hbv Infection ◽  
Luciferase Reporter ◽  
Artificial Transcription Factor ◽  
Chronic Hepatitis B Virus ◽  
B Virus

Chronic hepatitis B virus (HBV) infection is an independent risk factor for the development of hepatocellular carcinoma (HCC). The HBV HBx gene is frequently identified as an integrant in the chromosomal DNA of patients with HCC. HBx encodes the X protein (HBx), a putative viral oncoprotein that affects transcriptional regulation of several cellular genes. Therefore, HBx may be an ideal target to impede the progression of HBV infection–related HCC. In this study, integrated HBx was transcriptionally downregulated using an artificial transcription factor (ATF). Two three-fingered Cys2-His2 zinc finger (ZF) motifs that specifically recognized two 9-bp DNA sequences regulating HBx expression were identified from a phage-display library. The ZF domains were linked into a six-fingered protein that specified an 18-bp DNA target in the Enhancer I region upstream of HBx. This DNA-binding domain was fused with a Krüppel-associated box (KRAB) transcriptional repression domain to produce an ATF designed to downregulate HBx integrated into the Hep3B HCC cell line. The ATF significantly repressed HBx in a luciferase reporter assay. Stably expressing the ATF in Hep3B cells resulted in significant growth arrest, whereas stably expressing the ATF in an HCC cell line lacking integrated HBx (HepG2) had virtually no effect. The targeted downregulation of integrated HBx is a promising novel approach to inhibiting the progression of HBV infection–related HCC.

scholarly journals Production of hepatitis B virus in vitro by transient expression of cloned HBV DNA in a hepatoma cell line.

1987 ◽  
Vol 6 (3) ◽  
pp. 675-680 ◽  
Author(s):  
C.M. Chang ◽  
K.S. Jeng ◽  
C.P. Hu ◽  
S.J. Lo ◽  
T.S. Su ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Cell Line ◽  
Transient Expression ◽  
Hepatoma Cell ◽  
Hbv Dna ◽  
Hepatoma Cell Line ◽  
B Virus
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