Inhibitory Effect of TNF-α Produced by Macrophages Stimulated withGrifola frondosaExtract (ME) on the Growth of Influenza A/Aichi/2/68 Virus in MDCK Cells

2008 ◽  
Vol 36 (06) ◽  
pp. 1171-1183 ◽  
Author(s):  
Nobuko Obi ◽  
Katsumi Hayashi ◽  
Tatsurou Miyahara ◽  
Yutaka Shimada ◽  
Katsutoshi Terasawa ◽  
...  
Keyword(s):  
Influenza A ◽  
Mdck Cells ◽  
Inhibitory Effect ◽  
Fixed Dose ◽  
Madin Darby Canine Kidney ◽  
Tnf Α ◽  
Ultra Filtration ◽  
Infected Cells ◽  
Darby Canine Kidney

We investigated the inhibitory effect of the conditioned medium (CM) from P338D1 (D1) cells, a murine macrophage cell line, stimulated for 10 hours with a fixed dose (100 μg/ml) of the extracts from the fruit bodies of Grifola frondosa (ME) or its ultra filtration-based fractions (MFs), on the growth of influenza A/Aichi/2/68 virus in Madin-Darby canine kidney cells. Direct addition of ME and 3 kinds of MFs (MF1, MF2 and MF3) to the infected cells had no obvious inhibitory effect. However, virus yields were reduced in the presence of CMs. Notably, the inhibitory effect of the CM prepared by using MF2 (molecular weight of 30 Kd to 100 Kd) was the strongest (28% reduction compared to the control). RT-PCR and ELISA assays showed that the CMs could induce the expression of TNF-α mRNA in D1 cells leading to production of TNF-α, known as an antiviral cytokine. These findings suggest that ME and MFs (especially MF-2) might induce the production of certain factors, including TNF-α, which are responsible for the inhibition of viral growth in vitro.

Macrophage-Mediated Inhibitory Effect ofZingiber officinaleRosc, A Traditional Oriental Herbal Medicine, on the Growth of Influenza A/Aichi/2/68 Virus

2006 ◽  
Vol 34 (01) ◽  
pp. 157-169 ◽  
Author(s):  
Nobuko Imanishi ◽  
Tsugunobu Andoh ◽  
Naoki Mantani ◽  
Shinya Sakai ◽  
Katsutoshi Terasawa ◽  
...  
Keyword(s):  
Herbal Medicine ◽  
Influenza A ◽  
Mdck Cells ◽  
Zingiber Officinale ◽  
Inhibitory Effect ◽  
Aichi Virus ◽  
Tnf Α ◽  
Infected Cells ◽  
Necrosis Factor ◽  
Darby Canine Kidney

The inhibitory effect of Zingiber officinale Rosc (ZOR), an Oriental traditional herbal medicine, on the growth of influenza A/Aichi/2/68 (Aichi) virus was investigated in Madin-Darby canine kidney (MDCK) cells. Direct addition of ZOR (0.1 ~ 100 μ g/ml ) to the infected cells did not have any inhibitory effect. However, the ZOR-induced conditioned medium (ZOR-CM) of RAW cells, a murine macrophage (Mφ) cell line, exhibited an apparent inhibitory effect on MDCK cells without cytotoxicity. In accordance with the time-dependent inhibitory effect of ZOR-CM, it has been demonstrated that tumor necrosis factor (TNF)-α was gradually accumulated in ZOR-CM by the induction of TNF-α mRNA expression in ZOR-stimulated RAW cells. Conversely, the inhibitory effect of ZOR-CM was reduced significantly by the removal of TNF-α after the formation of an immune complex with anti-TNF-α monoclonal antibody. These data suggested that ZOR itself has no inhibitory effect on the growth of influenza virus, but could exert its effect via macrophage activation leading to production of TNF-α.

Nonmitogenic morphoregulatory action of pp60v-src on multicellular epithelial structures

1987 ◽  
Vol 7 (4) ◽  
pp. 1326-1337
Author(s):  
S L Warren ◽  
W J Nelson
Keyword(s):  
Tyrosine Kinases ◽  
Mdck Cells ◽  
Growth Potential ◽  
Junctional Complex ◽  
Madin Darby Canine Kidney ◽  
Epithelial Monolayers ◽  
Zonula Occludens ◽  
Darby Canine Kidney ◽  
Canine Kidney

Madin-Darby canine kidney (MDCK) cells form polarized, multicellular epithelial structures in vitro. Low-level expression of pp60v-src in MDCK cells elicits plasticity in these multicellular structures. Plasticity was revealed by the displacement of cells from mechanically stressed regions of the epithelial monolayers; however, the two-dimensional relationship between the cells in the remainder of the monolayer was maintained. Electron microscopy of multicellular structures revealed abnormal separation of the lateral membranes of adjacent cells and selective uncoupling of the junctional complex; the zonula adherens was disrupted, but the zonula occludens and desmosomes were retained. Significantly, this result was not accompanied by transformation of the cells, as judged by the absence of anchorage-independent growth potential. These results demonstrate a nonmitogenic biological activity of pp60v-src which is experimentally dissociable from transformation. This morphoregulatory action on higher-order epithelial structures may reflect a function of related cellular tyrosine kinases.

Transepithelial Resistance and Inulin Permeability as Endpoints in In Vitro Nephrotoxicity Testing

2002 ◽  
Vol 30 (2_suppl) ◽  
pp. 53-59 ◽  
Author(s):  
Tracey Duff ◽  
Simon Carter ◽  
Gemma Feldman ◽  
Gordon McEwan ◽  
Walter Pfaller ◽  
...  
Keyword(s):  
Proximal Tubule ◽  
Electrical Resistance ◽  
Mdck Cells ◽  
Ussing Chamber ◽  
Fluorescein Isothiocyanate ◽  
Transepithelial Electrical Resistance ◽  
Madin Darby Canine Kidney ◽  
Darby Canine Kidney ◽  
Canine Kidney

Transepithelial electrical resistance (RT) and the flux of fluorescein isothiocyanate (FITC) across Madin Darby canine kidney (MDCK) strain 1 cells and porcine epithelial kidney (LLC-PK1) monolayers were compared between three laboratories for a range of nephrotoxins. The precision of the REMS AutoSampler was similar to that of the Ussing chamber and the ENDOHM® technique, but superior to using chopstick electrodes, for measurements of resistance. The nephrotoxins used were selective for the proximal tubule, and in all cases, LLC-PK1 cells were more sensitive than MDCK cells. In most cases, change in RT was a more sensitive indicator of damage than alterations in FITC flux. The REMS system provides high intra-plate precision for RT measurements and is a higher throughput system, which is applicable to screening for nephrotoxicity in vitro.

scholarly journals Nonmitogenic morphoregulatory action of pp60v-src on multicellular epithelial structures.

1987 ◽  
Vol 7 (4) ◽  
pp. 1326-1337 ◽  
Author(s):  
S L Warren ◽  
W J Nelson
Keyword(s):  
Tyrosine Kinases ◽  
Mdck Cells ◽  
Growth Potential ◽  
Junctional Complex ◽  
Madin Darby Canine Kidney ◽  
Epithelial Monolayers ◽  
Zonula Occludens ◽  
Darby Canine Kidney ◽  
Canine Kidney

Madin-Darby canine kidney (MDCK) cells form polarized, multicellular epithelial structures in vitro. Low-level expression of pp60v-src in MDCK cells elicits plasticity in these multicellular structures. Plasticity was revealed by the displacement of cells from mechanically stressed regions of the epithelial monolayers; however, the two-dimensional relationship between the cells in the remainder of the monolayer was maintained. Electron microscopy of multicellular structures revealed abnormal separation of the lateral membranes of adjacent cells and selective uncoupling of the junctional complex; the zonula adherens was disrupted, but the zonula occludens and desmosomes were retained. Significantly, this result was not accompanied by transformation of the cells, as judged by the absence of anchorage-independent growth potential. These results demonstrate a nonmitogenic biological activity of pp60v-src which is experimentally dissociable from transformation. This morphoregulatory action on higher-order epithelial structures may reflect a function of related cellular tyrosine kinases.

scholarly journals Multigram Scale Synthesis and Anti-Influenza Activity of 3-Indoleacetonitrile Glucosides

2020 ◽  
Vol 15 (10) ◽  
pp. 1934578X2095328
Author(s):  
Jianghong Dong ◽  
Fengmei Yan ◽  
Shuang Li ◽  
Zewen Li ◽  
Yuanhang Qin
Keyword(s):  
Influenza A Virus ◽  
Influenza A ◽  
Phase Transfer ◽  
Madin Darby Canine Kidney ◽  
Influenza Activity ◽  
The Mannich Reaction ◽  
Key Steps ◽  
Glycosylation Reaction ◽  
Darby Canine Kidney

Indole-3-acetonitrile-6- O-β-d-glucopyranoside 1 is a simple alkaloid with anti-influenza A virus activity extracted from Radix isatidis. Herein, a concise and practical total synthetic route of 1 was presented, starting from 6-benzyloxyindole on a multigram scale. The pivotal reaction sequence involved the Mannich reaction, the fluoride ion-induced elimination-addition reaction, and phase-transfer-catalyzed glycosylation reaction as key steps. In addition, compounds 1 and two modified derivatives 8 and 16 were tested for anti-influenza efficacy and cytotoxicity in vitro performed on Madin-Darby canine kidney cells. The results revealed that the compounds exerted antiviral activity against the influenza A virus to a certain extent and displayed no cytotoxicity. These findings could contribute to the development of traditional Chinese medicine R. isatidis for treating the influenza virus.

scholarly journals Use of influenza A viruses expressing reporter genes to assess the frequency of double infections in vitro

2012 ◽  
Vol 93 (8) ◽  
pp. 1645-1648 ◽  
Author(s):  
R. Bodewes ◽  
N. J. Nieuwkoop ◽  
R. J. Verburgh ◽  
R. A. M. Fouchier ◽  
A. D. M. E. Osterhaus ◽  
...  
Keyword(s):  
Influenza A ◽  
Single Cells ◽  
Reporter Genes ◽  
Influenza Viruses ◽  
Influenza A Viruses ◽  
Double Infection ◽  
Madin Darby Canine Kidney ◽  
Darby Canine Kidney ◽  
Canine Kidney

Exchange of gene segments between mammalian and avian influenza A viruses may lead to the emergence of potential pandemic influenza viruses. Since co-infection of single cells with two viruses is a prerequisite for reassortment to take place, we assessed frequencies of double-infection in vitro using influenza A/H5N1 and A/H1N1 viruses expressing the reporter genes eGFP or mCherry. Double-infected A549 and Madin–Darby canine kidney cells were detected by confocal microscopy and flow cytometry.

scholarly journals Protective Role of Combined Polyphenols and Micronutrients against Influenza A Virus and SARS-CoV-2 Infection In Vitro

Biomedicines ◽  
2021 ◽  
Vol 9 (11) ◽  
pp. 1721
Author(s):  
Marta De Angelis ◽  
David Della-Morte ◽  
Gabriele Buttinelli ◽  
Angela Di Martino ◽  
Francesca Pacifici ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Influenza A ◽  
Viral Proteins ◽  
Antiviral Effect ◽  
Inhibitory Effect ◽  
Protective Role ◽  
Virus Infections ◽  
Infected Cells ◽  
Respiratory Virus Infections

Polyphenols have been widely studied for their antiviral effect against respiratory virus infections. Among these, resveratrol (RV) has been demonstrated to inhibit influenza virus replication and more recently, it has been tested together with pterostilbene against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. In the present work, we evaluated the antiviral activity of polydatin, an RV precursor, and a mixture of polyphenols and other micronutrients, named A5+, against influenza virus and SARS-CoV-2 infections. To this end, we infected Vero E6 cells and analyzed the replication of both respiratory viruses in terms of viral proteins synthesis and viral titration. We demonstrated that A5+ showed a higher efficacy in inhibiting both influenza virus and SARS-CoV-2 infections compared to polydatin treatment alone. Indeed, post infection treatment significantly decreased viral proteins expression and viral release, probably by interfering with any step of virus replicative cycle. Intriguingly, A5+ treatment strongly reduced IL-6 cytokine production in influenza virus-infected cells, suggesting its potential anti-inflammatory properties during the infection. Overall, these results demonstrate the synergic and innovative antiviral efficacy of A5+ mixture, although further studies are needed to clarify the mechanisms underlying its inhibitory effect.

scholarly journals STAT1 Is Required for Redifferentiation during Madin-Darby Canine Kidney Tubulogenesis

2010 ◽  
Vol 21 (22) ◽  
pp. 3926-3933 ◽  
Author(s):  
Minji Kim ◽  
Lucy Erin O'Brien ◽  
Sang-Ho Kwon ◽  
Keith E. Mostov
Keyword(s):  
Epithelial Mesenchymal Transition ◽  
Mdck Cells ◽  
Madin Darby Canine Kidney ◽  
Tubule Formation ◽  
Mesenchymal Transition ◽  
Phase Loss ◽  
Darby Canine Kidney ◽  
Canine Kidney ◽  
Constitutively Active

Tubule formation in vitro using Madin-Darby canine kidney (MDCK) epithelial cells consists mainly of two processes. First, the cells undergo a partial epithelial–mesenchymal transition (pEMT), losing polarity and migrating. Second, the cells redifferentiate, forming cords and then tubules with continuous lumens. We have shown previously that extracellular signal-regulated kinase activation is required for pEMT. However, the mechanism of how the pEMT phase is turned off and the redifferentiation phase is initiated is largely unknown. To address the central question of the sequential control of these two phases, we used MDCK cells grown as cysts and treated with hepatocyte growth factor to model tubulogenesis. We show that signal transducer and activator of transcription (STAT)1 controls the sequential progression from the pEMT phase to the redifferentiation phase. Loss of STAT1 prevents redifferentiation. Constitutively active STAT1 allows redifferentiation to occur even when cells are otherwise prevented from progressing beyond the pEMT phase by exogenous activation of Raf. Moreover, tyrosine phosphorylation defective STAT1 partially restored cord formation in such cells, suggesting that STAT1 functions in part as nonnuclear protein mediating signal transduction in this process. Constitutively active or inactive forms of STAT1 did not promote lumen maturation, suggesting this requires a distinct signal.

In-vitro anti Myxovirus Activity and Mechanism of Antiinfluenzavirus Activity of Polyoxometalates PM-504 and PM-523

1996 ◽  
Vol 7 (6) ◽  
pp. 346-352 ◽  
Author(s):  
S. Shigeta ◽  
S. Mori ◽  
J. Watanabe ◽  
T. Yamase ◽  
R. F. Schinazi
Keyword(s):  
Influenza A Virus ◽  
Influenza A ◽  
Mdck Cells ◽  
Cellular Membrane ◽  
Facs Analysis ◽  
Virus Envelope ◽  
Cytotoxic Compounds ◽  
Infected Cells ◽  
Chicken Erythrocytes

Sixty polyoxometalates were examined for anti-influenza A virus (FluV-A) activity in vitro. Two of the most potent and least cytotoxic compounds, PM-504 K9H5(Ge2Ti6W18O77)16H2O] and PM-523 (iPrNH3)6H [PTi2W10O38(O2)9H2O2] were selected for further studies. Examination of the antiviral effects of PM-504 and PM-523 against other human ortho- and paramyxoviruses revealed that both compounds had broad spectrum antimyxovirus activities. From a time of addition study and FACS analysis for influenza A virus infected cells, the compounds were found not to inhibit binding of virus to MDCK cells. However, these compounds inhibited haemolysis of chicken erythrocytes by virus and also inhibited fluorescence dequenching of octadecylrhodamine B-labelled virus after binding to cells. This indicates that these polyoxometalates inhibited fusion of the virus envelope to the cellular membrane.

An In Vitro Model of the Blood-Brain Barrier: The Response of Madin-Darby Canine Kidney Cells to Triethyl Tin

1996 ◽  
Vol 24 (3) ◽  
pp. 349-357
Author(s):  
Bellina Veronesi ◽  
Kent Carlsón ◽  
Marion Ehrich
Keyword(s):  
Cell Line ◽  
Blood Brain Barrier ◽  
Electrical Resistance ◽  
Mdck Cells ◽  
Brain Barrier ◽  
Madin Darby Canine Kidney ◽  
Blood Brain ◽  
Darby Canine Kidney ◽  
Canine Kidney

The development of a cell culture model which simulates the properties of the blood–brain barrier (BBB) is necessary for the detection of neurotoxic chemicals that can disrupt the barrier, and to provide a more “risk relevant” in vitro screening battery. The present study evaluates the Madin-Darby canine kidney (MDCK) epithelial cell line for this purpose. Changes in electrical resistance and enzyme activities were correlated in confluent MDCK cells exposed to the neurotoxic metal, triethyl tin (TET). Concentrations of TET (0.001–10μM) were established that produced depression in electrical resistance of the MDCK cells after exposure for 8 hours or caused fluorescein leakage after exposure for 72 hours. Confluent cultures of MDCK cells were then exposed to these concentrations of TET and assayed after exposure for 24 hours and 72 hours for changes in those enzymes common to both epithelial and cerebral endothelial cells. The results indicated that increased alkaline phosphatase (APP), γ-glutamyl transpeptidase (GGTP) and superoxide dismutase (SOD) characterised the loss of electrical resistance and permeability disruption in TET-exposed MDCK confluent cultures. Relative increases in APP and decreases in GGTP activities preceded cytotoxicity, which was associated with a high SOD activity. Such enzyme changes may be predictive endpoints of barrier cell disruption by neurotoxic metals in this cell line and support the additional evaluation of the MDCK cell line as an in vitro “screen” for chemicals that disrupt the BBB.

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