Comparative characterization of flavivirus production in two cell lines: Human hepatoma-derived Huh7.5.1-8 and African green monkey kidney-derived Vero

Saito, Kyoko; Fukasawa, Masayoshi; Shirasago, Yoshitaka; Suzuki, Ritsuro; Osada, Naoki; Yamaji, Taiki; Wakita, Takaji; Konishi, Eiji; Hanada, Kentaro

doi:10.1371/journal.pone.0232274

Cited by 9 publications

(7 citation statements)

References 30 publications

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“…For instance, C6/36 cells exhibit altered RNA-mediated antiviral immunity ( Brackney et al, 2010 ; Scott et al, 2010 ). Huh7.5.1 cells are RIG-I-deficient ( Shirasago et al, 2015 ; Zhong et al, 2006 ) while Vero cells are deficient in type-I interferon production ( Saito et al, 2020 ; Desmyter et al, 1968 ; Osada et al, 2014 ). Intriguingly, viral populations exhibit an intermediate phenotype in Huh7.5.1 cells relative to the other primate and insect cell lines, which are distinct from their phenotype in Vero cells.…”

Section: Resultsmentioning

confidence: 99%

Principles of dengue virus evolvability derived from genotype-fitness maps in human and mosquito cells

Dolan

Taguwa

Rangel

et al. 2021

eLife

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Dengue virus (DENV) cycles between mosquito and mammalian hosts. To examine how DENV populations adapt to these different host environments we used serial passage in human and mosquito cell lines and estimated fitness effects for all single-nucleotide variants in these populations using ultra-deep sequencing. This allowed us to determine the contributions of beneficial and deleterious mutations to the collective fitness of the population. Our analysis revealed that the continuous influx of a large burden of deleterious mutations counterbalances the effect of rare, host-specific beneficial mutations to shape the path of adaptation. Beneficial mutations preferentially map to intrinsically disordered domains in the viral proteome and cluster to defined regions in the genome. These phenotypically redundant adaptive alleles may facilitate host-specific DENV adaptation. Importantly, the evolutionary constraints described in our simple system mirror trends observed across DENV and Zika strains, indicating it recapitulates key biophysical and biological constraints shaping long-term viral evolution.

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Section: Resultsmentioning

confidence: 99%

Principles of dengue virus evolvability derived from genotype-fitness maps in human and mosquito cells

Dolan

Taguwa

Rangel

et al. 2021

eLife

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“…It was investigated by plaque inhibition assay after the determination of its cytotoxic activity on Vero-E6 cells. These cells are derived from the kidney of an African green monkey and they are widely used in virology studies [28]. APEE exhibited potent antiviral activity against SARS-CoV-2 in Vero-E6 cells with a value of IC 50 1.1 ± 0.03 µg/mL.…”

Section: Discussionmentioning

confidence: 99%

Promising Antiviral Activity of Agrimonia pilosa Phytochemicals against Severe Acute Respiratory Syndrome Coronavirus 2 Supported with In Vivo Mice Study

et al. 2021

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The global emergence of the COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has focused the entire world’s attention toward searching for a potential remedy for this disease. Thus, we investigated the antiviral activity of Agrimonia pilosa ethanol extract (APEE) against SARS-CoV-2 and it exhibited a potent antiviral activity with IC50 of 1.1 ± 0.03 µg/mL. Its mechanism of action was elucidated, and it exhibited a virucidal activity and an inhibition of viral adsorption. Moreover, it presented an immunomodulatory activity as it decreased the upregulation of gene expression of COX-2, iNOS, IL-6, TNF-α, and NF-κB in lipopolysaccharide (LPS)-induced peripheral blood mononuclear cells. A comprehensive analysis of the phytochemical fingerprint of APEE was conducted using LC-ESI-MS/MS technique for the first time. We detected 81 compounds and most of them belong to the flavonoid and coumarin classes. Interestingly, isoflavonoids, procyanidins, and anthocyanins were detected for the first time in A. pilosa. Moreover, the antioxidant activity was evidenced in DPPH (IC50 62.80 µg/mL) and ABTS (201.49 mg Trolox equivalents (TE)/mg) radical scavenging, FRAP (60.84 mg TE/mg), and ORAC (306.54 mg TE/g) assays. Furthermore, the protective effect of APEE was investigated in Lipopolysaccharides (LPS)-induced acute lung injury (ALI) in mice. Lung W/D ratio, serum IL-6, IL-18, IL-1β, HO-1, Caspase-1, caspase-3, TLR-4 expression, TAC, NO, MPO activity, and histopathological examination of lung tissues were assessed. APEE induced a marked downregulation in all inflammation, oxidative stress, apoptosis markers, and TLR-4 expression. In addition, it alleviated all histopathological abnormalities confirming the beneficial effects of APEE in ALI. Therefore, APEE could be a potential source for therapeutic compounds that could be investigated, in future preclinical and clinical trials, in the treatment of patients with COVID-19.

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“…Therefore, we resequenced the genomic DNA from Huh7 and Huh7.5.1-8 using a Sanger sequencer and verified that the mutation is absent in Huh7 but heterozygous in Huh7.5.1-8 ( Figure 2). Our recent study showed that a 55T allele transcript harbors an allele-specific large deletion and, presumably, nonfunctional in Huh7.5.1-8 (Saito et al, 2020). The results showed that, although T55I mutation is heterozygous in the Huh7.5.1-8 genome, all full-length RIG-I proteins have the T55I mutation.…”

Section: Missense Mutation In Rig-i (T55i) Wasmentioning

confidence: 97%

Identification of Characteristic Genomic Markers in Human Hepatoma HuH-7 and Huh7.5.1-8 Cell Lines

et al. 2020

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The human hepatoma-derived HuH-7 cell line and its derivatives (Huh7.5 and Huh7.5.1) have been widely used as a convenient experimental substitute for primary hepatocytes. In particular, these cell lines represent host cells suitable for propagating the hepatitis C virus (HCV) in vitro. The Huh7.5.1-8 cell line, a subline of Huh7.5.1, can propagate HCV more efficiently than its parental cells. To provide genomic information for cells' quality control, we performed whole-genome sequencing of HuH-7 and Huh7.5.1-8 and identified their characteristic genomic deletions, some of which are applicable to an in-house test for cell authentication. Among the genes related to HCV infection and replication, 53 genes were found to carry missense or loss-of-function mutations likely specific to the HuH-7 and/or Huh7.5.1-8. Eight genes, including DDX58 (RIG-I), BAX, EP300, and SPP1 (osteopontin), contained mutations observed only in Huh7.5.1-8 or mutations with higher frequency in Huh7.5.1-8. These mutations might be relevant to phenotypic differences between the two cell lines and may also serve as genetic markers to distinguish Huh7.5.1-8 cells from the ancestral HuH-7 cells.

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Comparative characterization of flavivirus production in two cell lines: Human hepatoma-derived Huh7.5.1-8 and African green monkey kidney-derived Vero

Cited by 9 publications

References 30 publications

Principles of dengue virus evolvability derived from genotype-fitness maps in human and mosquito cells

Principles of dengue virus evolvability derived from genotype-fitness maps in human and mosquito cells

Promising Antiviral Activity of Agrimonia pilosa Phytochemicals against Severe Acute Respiratory Syndrome Coronavirus 2 Supported with In Vivo Mice Study

Identification of Characteristic Genomic Markers in Human Hepatoma HuH-7 and Huh7.5.1-8 Cell Lines

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