Microculture virus titration — a simple colourimetric assay for influenza virus titration

Levi, Raphael; Beeor-Tzahar, Talia; Arnon, Ruth

doi:10.1016/0166-0934(94)00137-6

Cited by 22 publications

(12 citation statements)

References 11 publications

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“…A colorimetric MTT assay was performed as previously described (47). Color adsorption in the solution was measured using a microplate reader (BioTek EL 800; BioTek Instruments Inc., Winooski, VT, USA) at 570 nm.…”

Section: Methodsmentioning

confidence: 99%

Simvastatin modulates cellular components in influenza A virus-infected cells

Mehrbod

Hair-Bejo

Ibrahim

et al. 2014

International Journal of Molecular Medicine

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Influenza A virus is one of the most important health risks that lead to significant respiratory infections. Continuous antigenic changes and lack of promising vaccines are the reasons for the unsuccessful treatment of influenza. Statins are pleiotropic drugs that have recently served as anti-influenza agents due to their anti-inflammatory activity. In this study, the effect of simvastatin on influenza A-infected cells was investigated. Based on the MTT cytotoxicity test, hemagglutination (HA) assay and qPCR it was found that simvastatin maintained cell viability and decreased the viral load significantly as compared to virus-inoculated cells. The expression of important pro-inflammatory cytokines (tumor necrosis factor-α, interleukin-6 and interferon-γ), which was quantified using ELISA showed that simvastatin decreased the expression of pro-inflammatory cytokines to an average of 2-fold. Furthermore, the modulation of actin filament polymerization was determined using rhodamine staining. Endocytosis and autophagy processes were examined by detecting Rab and RhoA GTPase protein prenylation and LC3 lipidation using western blotting. The results showed that inhibiting GTPase and LC3 membrane localization using simvastatin inhibits influenza replication. Findings of this study provide evidence that modulation of RhoA, Rabs and LC3 may be the underlying mechanisms for the inhibitory effects of simvastatin as an anti-influenza compound.

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Section: Methodsmentioning

confidence: 99%

Simvastatin modulates cellular components in influenza A virus-infected cells

Mehrbod

Hair-Bejo

Ibrahim

et al. 2014

International Journal of Molecular Medicine

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“…Using a well-established in-vitro model of influenza A virus infection (9,40,50,52), we investigated the impact of pH1N1 virus infection of human respiratory epithelial cells by comparing three pH1N1 viruses (2,4,14), evaluating virus infectivity and growth, the early host antiviral and inflammatory responses (31,33,61), and virus-induced cell death (13,39,40). Our results show that compared with a recent seasonal vaccine strain of H1N1 virus (A/Solomon Islands/ 3/2006, hereafter referred to as A/Sol), and more importantly among themselves, pH1N1 viruses demonstrate significant differences in early antiviral cytokine and inflammatory responses, as well as cell death.…”

Section: Introductionmentioning

confidence: 99%

Infection of Lung Epithelial Cells with Pandemic 2009 A(H1N1) Influenza Viruses Reveals Isolate-Specific Differences in Infectivity and Host Cellular Responses

et al. 2011

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To better understand the early virus-host interactions of the pandemic 2009 A(H1N1) viruses in humans, we examined early host responses following infection of human epithelial cell cultures with three 2009 A(H1N1) viruses (A/California/08/2009, A/Mexico/4108/2009, and A/Texas/15/2009), or a seasonal H1N1 vaccine strain (A/Solomon Islands/3/2006). We report here that infection with pandemic A/California/08/2009 and A/Mexico/4108/2009 viruses resulted in differences in virus infectivity compared to either pandemic A/Texas/15/2009 or the seasonal H1N1 vaccine strain. In addition, IFN-β levels were decreased in cell cultures infected with either the A/California/08/2009 or the A/Mexico/4108/2009 virus. Furthermore, infection with A/California/08/2009 and A/Mexico/4108/2009 viruses resulted in lower expression of four key proinflammatory markers (IL-6, RANTES, IP-10, and MIP-1β) compared with infection with either A/Texas/15/2009 or A/Solomon Islands/3/2006. Taken together, our results demonstrate that 2009 A(H1N1) viruses isolated during the Spring wave induced varying degrees of early host antiviral and inflammatory responses in human respiratory epithelial cells, highlighting the strain-specific nature of these responses, which play a role in clinical disease.

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“…Virus growth and purification were performed according to standard methods as described by Barret and Inglis (30). The titer of virus used for infection was evaluated by the infection of Madin-Darby canine kidney (MDCK) cells (31), and hence, virus titer was expressed as the tissue culture infective doses leading to 50% infected cells (TCID50).…”

Section: Methodsmentioning

confidence: 99%

A DNA Aptamer Prevents Influenza Infection by Blocking the Receptor Binding Region of the Viral Hemagglutinin

Jeon¹,

Kayhan²,

Ben-Yedidia³

et al. 2004

Journal of Biological Chemistry

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157

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Influenza A virus infection is a major source of morbidity and mortality worldwide. Current means of control for influenza are based on prophylaxis by vaccines and on treatment by the available specific influenza neuraminidase inhibitor drugs. The approach taken in the present study is to prevent and/or ameliorate influenza infection by site-specific blocking of the viral binding to host cell receptors. We describe a novel oligonucleotide, known also as an aptamer, which has been designed to complement the receptor-binding region of the influenza hemagglutinin molecule. It was constructed by screening a DNA library and processing by the selective evolution of ligands by exponential enrichment (SELEX) procedure. We show that this DNA aptamer is indeed capable of inhibiting the hemagglutinin capacity of the virus, as well as in the prevention of viral infectivity in vitro, in tissue culture. Furthermore, it inhibits viral infection by different influenza strains in an animal model, as manifested by 90 -99% reduction of virus burden in the lungs of treated mice. The mode of action of this aptamer is by blocking the binding of influenza virus to target cell receptors and consequently prevention of the virus invasion into the host cells.

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Microculture virus titration — a simple colourimetric assay for influenza virus titration

Cited by 22 publications

References 11 publications

Simvastatin modulates cellular components in influenza A virus-infected cells

Simvastatin modulates cellular components in influenza A virus-infected cells

Infection of Lung Epithelial Cells with Pandemic 2009 A(H1N1) Influenza Viruses Reveals Isolate-Specific Differences in Infectivity and Host Cellular Responses

A DNA Aptamer Prevents Influenza Infection by Blocking the Receptor Binding Region of the Viral Hemagglutinin

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