scholarly journals Antiviral Evaluation of UV-4B and Interferon-Alpha Combination Regimens against Dengue Virus

Viruses ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 771
Author(s):  
Evelyn J. Franco ◽  
Camilly P. Pires de Mello ◽  
Ashley N. Brown
Keyword(s):  
Dengue Virus ◽  
Cell Lines ◽  
Interferon Alpha ◽  
Plaque Assay ◽  
Clinical Manifestations ◽  
Cell Types ◽  
Vero Cells ◽  
Human Infection ◽  
Severe Dengue ◽  
Viral Burden

Dengue virus (DENV) is a flavivirus associated with clinical manifestations ranging in severity from self-limiting dengue fever, to the potentially life threatening condition, severe dengue. There are currently no approved antiviral therapies for the treatment of DENV. Here, we evaluated the antiviral potential of four broad-spectrum antivirals, UV-4B, interferon-alpha (IFN), sofosbuvir (SOF), and favipiravir (FAV) against DENV serotype 2 as mono- and combination therapy in cell lines that are physiologically relevant to human infection. Cell lines derived from human liver (HUH-7), neurons (SK-N-MC), and skin (HFF-1) were infected with DENV and treated with UV-4B, IFN, SOF, or FAV. Viral supernatant was sampled daily and infectious viral burden was quantified by plaque assay on Vero cells. Drug effect on cell proliferation in uninfected and infected cells was also assessed. UV-4B inhibited DENV in HUH-7, SK-N-MC, and HFF-1 cells yielding EC50 values of 23.75, 49.44, and 37.38 µM, respectively. Clinically achievable IFN concentrations substantially reduced viral burden in HUH-7 (EC50 = 102.7 IU/mL), SK-N-MC (EC50 = 86.59 IU/mL), and HFF-1 (EC50 = 163.1 IU/mL) cells. SOF potently inhibited DENV in HUH-7 cells but failed to produce the same effect in SK-N-MC and HFF-1 cells. Finally, FAV provided minimal suppression in HUH-7 and SK-N-MC cells, but was ineffective in HFF-1 cells. The two most potent anti-DENV agents, UV-4B and IFN, were also assessed in combination. UV-4B + IFN treatment enhanced antiviral activity in HUH-7, SK-N-MC, and HFF-1 cells relative to monotherapy. Our results demonstrate the antiviral potential of UV-4B and IFN against DENV in multiple physiologically relevant cell types.

scholarly journals Combination Regimens of Favipiravir Plus Interferon Alpha Inhibit Chikungunya Virus Replication in Clinically Relevant Human Cell Lines

2021 ◽  
Vol 9 (2) ◽  
pp. 307
Author(s):  
Evelyn J. Franco ◽  
Xun Tao ◽  
Kaley C. Hanrahan ◽  
Jieqiang Zhou ◽  
Jürgen B. Bulitta ◽  
...  
Keyword(s):  
Combination Therapy ◽  
Antiviral Activity ◽  
Viral Replication ◽  
Cell Lines ◽  
Interferon Alpha ◽  
Chikungunya Virus ◽  
Plaque Assay ◽  
Human Infection ◽  
Viral Burden ◽  
Combination Regimens

Chikungunya virus (CHIKV) is an alphavirus associated with a broad tissue tropism for which no antivirals or vaccines are approved. This study evaluated the antiviral potential of favipiravir (FAV), interferon-alpha (IFN), and ribavirin (RBV) against CHIKV as mono- and combination-therapy in cell lines that are clinically relevant to human infection. Cells derived from human connective tissue (HT-1080), neurons (SK-N-MC), and skin (HFF-1) were infected with CHIKV and treated with different concentrations of FAV, IFN, or RBV. Viral supernatant was sampled daily and the burden was quantified by plaque assay on Vero cells. FAV and IFN were the most effective against CHIKV on various cell lines, suppressing the viral burden at clinically achievable concentrations; although the degree of antiviral activity was heavily influenced by cell type. RBV was not effective and demonstrated substantial toxicity, indicating that it is not a feasible candidate for CHIKV. The combination of FAV and IFN was then assessed on all cell lines. Combination therapy enhanced antiviral activity in HT-1080 and SK-N-MC cells, but not in HFF-1 cells. We developed a pharmacokinetic/pharmacodynamic model that described the viral burden and inhibitory antiviral effect. Simulations from this model predicted clinically relevant concentrations of FAV plus IFN completely suppressed CHIKV replication in HT-1080 cells, and considerably slowed down the rate of viral replication in SK-N-MC cells. The model predicted substantial inhibition of viral replication by clinical IFN regimens in HFF-1 cells. Our results highlight the antiviral potential of FAV and IFN combination regimens against CHIKV in clinically relevant cell types.

scholarly journals Zika Virus Replication Is Substantially Inhibited by Novel Favipiravir and Interferon Alpha Combination Regimens

2017 ◽  
Vol 62 (1) ◽  
Author(s):  
Camilly P. Pires de Mello ◽  
Xun Tao ◽  
Tae Hwan Kim ◽  
Jürgen B. Bulitta ◽  
Jaime L. Rodriquez ◽  
...  
Keyword(s):  
Interferon Alpha ◽  
Zika Virus ◽  
Time Course ◽  
Plaque Assay ◽  
Vero Cells ◽  
Health Concern ◽  
Combination Regimen ◽  
Viral Inhibition ◽  
Dosage Regimens ◽  
Viral Burden

ABSTRACT Zika virus (ZIKV) is a major public health concern due to its overwhelming spread into the Americas. Currently, there are neither licensed vaccines nor antiviral therapies available for the treatment of ZIKV. We aimed to identify and rationally optimize effective therapeutic regimens for ZIKV by evaluating the antiviral potentials of the approved broad-spectrum antiviral agents favipiravir (FAV), interferon alpha (IFN), and ribavirin (RBV) as single agents and in combinations. For these studies, Vero cells were infected with ZIKV in the presence of increasing concentrations of FAV, IFN, or/and RBV for 4 days. Supernatants were harvested daily, and the viral burden was quantified by a plaque assay on Vero cells. The time course of the viral burden during treatment in vitro was characterized by a novel translational, mechanism-based model, which was subsequently used to rationally optimize combination dosage regimens. The combination regimen of FAV plus IFN provided the greatest extent of viral inhibition without cytotoxicity, reducing the viral burden by 4.4 log10 PFU/ml at concentrations of 250 μM FAV and 100 IU/ml IFN. Importantly, these concentrations are achievable in humans. The translational, mechanism-based model yielded unbiased and reasonably precise curve fits. Simulations with the model predicted that clinically relevant regimens of FAV plus IFN would markedly reduce viral burdens in humans, resulting in at least a 10,000-fold reduction in the amount of the virus during the first 4 days of treatment. These findings highlight the substantial promise of rationally optimized combination dosage regimens of FAV plus IFN, which should be further investigated to combat ZIKV.

scholarly journals The effectiveness of antiviral agents with broad-spectrum activity against chikungunya virus varies between host cell lines

2018 ◽  
Vol 26 ◽  
pp. 204020661880758 ◽  
Author(s):  
Evelyn J Franco ◽  
Jaime L Rodriquez ◽  
Justin J Pomeroy ◽  
Kaley C Hanrahan ◽  
Ashley N Brown
Keyword(s):  
Antiviral Activity ◽  
Host Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Broad Spectrum ◽  
Chikungunya Virus ◽  
Plaque Assay ◽  
A549 Cells ◽  
Western Hemisphere ◽  
Viral Burden

Chikungunya virus (CHIKV) is a mosquito-borne virus that has recently emerged in the Western Hemisphere. Approved antiviral therapies or vaccines for the treatment or prevention of CHIKV infections are not available. This study aims to evaluate the antiviral activity of commercially available broad-spectrum antivirals against CHIKV. Due to host cell-specific variability in uptake and intracellular processing of drug, we evaluated the antiviral effects of each agent in three cell lines. Antiviral activities of ribavirin (RBV), interferon-alfa (IFN-α) and favipiravir (FAV) were assessed in CHIKV-infected Vero, HUH-7, and A549 cells. CHIKV-infected cells were treated with increasing concentrations of each agent for three days and viral burden was quantified by plaque assay on Vero cells. Cytotoxic effects of RBV, FAV and IFN-α were also evaluated. Antiviral activity differed depending on the cell line used for evaluation. RBV had the greatest antiviral effect in HUH-7 cells (EC50 = 2.575 µg/mL); IFN-α was most effective in A549 cells (EC50 = 4.235 IU/mL); and FAV in HUH-7 cells (EC50 = 20.00 μg/mL). The results of our study show FAV and IFN-α are the most promising candidates, as their use led to substantial reductions in viral burden at clinically achievable concentrations in two human-derived cell lines. FAV is an especially attractive candidate for further investigation due to its oral bioavailability. These findings also highlight the importance of cell line selection for preclinical drug trials.

scholarly journals EvaluatingShigella flexneriPathogenesis in the Human Enteroid Model

2019 ◽  
Vol 87 (4) ◽  
Author(s):  
Sridevi Ranganathan ◽  
Michele Doucet ◽  
Christen L. Grassel ◽  
BreOnna Delaine-Elias ◽  
Nicholas C. Zachos ◽  
...  
Keyword(s):  
Animal Models ◽  
Cell Lines ◽  
System Modeling ◽  
Human Colon ◽  
Cell Types ◽  
Human Infection ◽  
Content Type ◽  
Transformed Cell ◽  
Severe Diarrhea ◽  
Transformed Cell Lines

ABSTRACTThe enteric pathogenShigellais one of the leading causes of moderate-to-severe diarrhea and death in young children in developing countries. Transformed cell lines and animal models have been widely used to studyShigellapathogenesis. In addition to altered physiology, transformed cell lines are composed of a single cell type that does not sufficiently represent the complex multicellular environment of the human colon. Most available animal models do not accurately mimic human disease. The human intestinal enteroid model, derived from LGR5+stem cell-containing intestinal crypts from healthy subjects, represents a technological leap in human gastrointestinal system modeling and provides a more physiologically relevant system that includes multiple cell types and features of the human intestine. We established the utility of this model for studying basic aspects ofShigellapathogenesis and host responses. In this study, we show thatShigellaflexneriis capable of infecting and replicating intracellularly in human enteroids derived from different segments of the intestine. Apical invasion byS. flexneriis very limited but increases ∼10-fold when enteroids are differentiated to include M cells. Invasion via the basolateral surface was at least 2-log10units more efficient than apical infection. Increased secretion of interleukin-8 and higher expression levels of the mucin glycoprotein Muc2 were observed in the enteroids followingS. flexneriinfection. The human enteroid model promises to bridge some of the gaps between traditional cell culture, animal models, and human infection.

scholarly journals Attachment and invasion ofToxoplasma gondiiandNeospora caninumto epithelial and fibroblast cell linesin vitro

Parasitology ◽  
2005 ◽  
Vol 131 (5) ◽  
pp. 583-590 ◽  
Author(s):  
YING LEI ◽  
M. DAVEY ◽  
J. T. ELLIS
Keyword(s):  
Cell Lines ◽  
Neospora Caninum ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Vero Cells ◽  
Host Cells ◽  
Trypsin Treatment ◽  
Epithelial Cell Lines ◽  
Pre Treatment

Attachment and invasion ofToxoplasma gondiiandNeospora caninumto a cat and a dog fibroblast cell line and 2 epithelial cell lines (a cat kidney and Vero) were comparedin vitrousing fluorescence antibody methodology. In addition, trypsin treatment of tachyzoites was used to determine whether protein molecules were essential to the process of invasion. The results show that bothT. gondiiandN. caninuminvaded all 4 cell lines, and that pre-treatment ofT. gondiitachyzoites with trypsin caused an increase in the ability of the parasite to invade these host cells. FurthermoreT. gondii, in comparison toN. caninum, invaded all 4 cell lines at greater levels. The results here support the conclusion that bothT. gondiiandN. caninumhave the ability to invade a variety of cell types including both dog and cat cells, and questions the utility of Vero cells as an appropriate host cell forin vitrostudies on the biology of these taxa.

scholarly journals Chloroquine Inhibits Dengue Virus Type 2 Replication in Vero Cells but Not in C6/36 Cells

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Kleber Juvenal Silva Farias ◽  
Paula Renata Lima Machado ◽  
Benedito Antônio Lopes da Fonseca
Keyword(s):  
Dengue Virus ◽  
Virus Type ◽  
Mammalian Cells ◽  
Plaque Assay ◽  
Antiviral Drug ◽  
Vero Cells ◽  
Significant Difference ◽  
Dengue Virus Type 2

Dengue viruses are the most important arthropod-borne viruses in terms of morbidity and mortality in the world. Since there is no dengue vaccine available for human use, we have set out to investigate the use of chloroquine as an antiviral drug against dengue. Chloroquine, an amine acidotropic drug known to affect intracellular exocytic pathways by increasing endosomal pH, was used in the in vitro treatment of Vero and C6/36 cells infected with dengue virus type 2 (DENV-2). Real-time RT-PCR and plaque assays were used to quantify the DENV-2 load in infected Vero and C6/36 cells after chloroquine treatment. Our results showed that a dose of 50 μg/ml of chloroquine was not toxic to the cells and induced a statistically significant inhibition of virus production in infected Vero cells when compared to untreated cells. In C6/36 cells, chloroquine does not induce a statistically significant difference in viral replication when compared to untreated cells, showing that this virus uses an unlikely pathway of penetration in these cells, and results were also confirmed by the plaque assay (PFU). These data suggest that the inhibition of virus infection induced by chloroquine is due to interference with acidic vesicles in mammalian cells.

Dengue virus binding to human leukocyte cell lines: receptor usage differs between cell types and virus strains

2001 ◽  
Vol 73 (1) ◽  
pp. 81-89 ◽  
Author(s):  
Helle Bielefeldt-Ohmann ◽  
Michelle Meyer ◽  
David R Fitzpatrick ◽  
John S Mackenzie
Keyword(s):  
Dengue Virus ◽  
Cell Lines ◽  
Human Leukocyte ◽  
Cell Types ◽  
Virus Binding ◽  
Receptor Usage ◽  
Virus Strains

scholarly journals Karakteristik Klinis dan Virologis Penderita Demam Berdarah Dengue di Kota Bandar Lampung

2020 ◽  
Vol 12 (2) ◽  
pp. 85-92
Author(s):  
Nurminha Nurminha ◽  
Tori Rihiantoro ◽  
Mara Ipa
Keyword(s):  
Dengue Virus ◽  
Clinical Symptoms ◽  
Serological Test ◽  
Secondary Infection ◽  
Dengue Infection ◽  
Clinical Manifestations ◽  
Denv Infection ◽  
Severe Dengue ◽  
Virus Serotype ◽  
Degree Of Severity

Abstract. Clinical symptoms of dengue virus (DENV) infection range from asymptomatic mild dengue fever(DF), more severe dengue hemorrhagic fever (DHF) up to dengue shock syndrome. One of the determinantsof dengue infection severity was virus virulence. This study aimed to determine the clinical and virologicalcharacteristics of dengue virus infection patients based on the severity degree. A cross-sectional study wasconducted in RSUD Dr. H. Abdul Moeloek, Lampung Province between July-November 2016 with 56 denguepatients as samples selected using purposive sampling. The serological test was done using a rapiddiagnostic test. Blood samples for DENV serotype identification were examined using reverse-transcriptionpolymerase chain reaction. Classification of DENV infection severity was obtained from the patient’s medicalrecord. The results showed that the most common clinical manifestations were fever, headache, and retroorbitalpain, appearing in all patients from every degree of severity. There were Grade I DHF patients whoexperienced Myalgia (15.6%) and petechiae (22.2%). Laboratory results showed that thrombocytopeniaappeared in every grade, even though 13.3% of grade I patients did not experience it. Secondary infectionwas found in 92.9% of samples, and all DENV serotype can be detected in 39.2%samples: DENV-1 (46.7%),DENV-2 (6.7%), DENV-3 (26.7%), and DENV-4 (20%). This study concluded that the majority of clinicalcharacteristics in DHF patients are in line with the degree of severity, with the bleeding as the dominantmanifestation in patients with grade II-IV. Virological characteristics of DENV-1 are dominant in all patientswith DHF grade I-IV.Keywords: dengue virus, serotype, severity, secondary infection, Bandar Lampung

scholarly journals Differential inhibition of dengue virus infection in mammalian and mosquito cells by iota-carrageenan

2011 ◽  
Vol 92 (6) ◽  
pp. 1332-1342 ◽  
Author(s):  
Laura B. Talarico ◽  
Miguel D. Noseda ◽  
Diogo R. B. Ducatti ◽  
Maria E. R. Duarte ◽  
Elsa B. Damonte
Keyword(s):  
Dengue Virus ◽  
Mammalian Cells ◽  
Differential Susceptibility ◽  
Cell Types ◽  
Vero Cells ◽  
Yield Reduction ◽  
Structural Class ◽  
Mosquito Cells ◽  
Refractory State ◽  
Pre Treatment

The antiviral activity against dengue virus-2 (DENV-2) of carrageenans reported here has shown a differential susceptibility of C6/36 HT and Vero cells, taken as models of mosquito and mammalian cells, depending on the structural class of polysaccharides: all polysaccharides blocked DENV-2 infection in monkey Vero cells, but only iota-carrageenans were virus inhibitors in mosquito cells. However, iota-carrageenans were less effective in mosquito cells in comparison with mammalian cells with effective concentration 50 % (EC50) values in C6/36 HT cells 4.9–17.5-fold higher than in Vero cells, as determined by virus yield reduction assay. The mode of action of iota-carrageenan in both cell types was strikingly different: in Vero cells the inhibitory activity was exerted only at the initiation of the cycle, affecting virion binding, whereas in mosquito cells DENV-2 adsorption was not affected and comparable levels of inhibition were obtained if the compound was added to cells together with the virus, after 8 h of infection or by cell pre-treatment before infection. Furthermore, iota-carrageenans induced a subtle alteration in mosquito cells, detected by cell proliferation and protein synthesis analyses, suggesting that a probable cellular target may be responsible for the refractory state of mosquito cells to DENV-2 infection produced by this class of polysulfates. The failure of iota-carrageenan to block DENV-2 adsorption to mosquito cells appeared to be related to the low presence of adequate heparan sulfate (HS) in C6/36 HT cell surface and is indicative of a differential participation of HS residues for DENV-2 entry in both types of cells.

scholarly journals Modulation of Dengue Virus Infection in Human Cells by Alpha, Beta, and Gamma Interferons

2000 ◽  
Vol 74 (11) ◽  
pp. 4957-4966 ◽  
Author(s):  
Michael S. Diamond ◽  
T. Guy Roberts ◽  
Dianna Edgil ◽  
Betty Lu ◽  
Joel Ernst ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Plaque Assay ◽  
Cell Types ◽  
Viral Rna ◽  
Rt Pcr ◽  
Cell Infection ◽  
Variable Effect ◽  
Dependent Cell ◽  
Low Passage

ABSTRACT A role for interferon (IFN) in modulating infection by dengue virus (DV) has been suggested by studies in DV-infected patients and IFN receptor-deficient mice. To address how IFN modulates DV type 2 infection, we have assayed IFN-α, -β, and -γ for the ability to enhance or diminish antibody-independent and antibody-dependent cell infection using a competitive, asymmetric reverse transcriptase-mediated PCR (RT-PCR) assay that quantitates positive and negative strands of viral RNA, a flow cytometric assay that measures viral antigen, and a plaque assay that analyzes virion production. Our data suggest that IFN-α and -β protect cells against DV infection in vitro. Treatment of hepatoma cells with IFN-α or -β decreases viral RNA levels greater than 1,000-fold, the percentage of cells infected 90 to 95%, and the amount of infectious virus secreted 150- to 100,000-fold. These results have been reproduced with several cell types and viral strains, including low-passage isolates. In contrast, IFN-γ has a more variable effect depending on the cell type and pathway of infection. Quantitative RT-PCR experiments indicate that IFN inhibits DV infection by preventing the accumulation of negative-strand viral RNA.

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