Polarity of Human Parainfluenza Virus Type 3 Infection in Polarized Human Lung Epithelial A549 Cells: Role of Microfilament and Microtubule

Bose, Santanu; Malur, Anagha; Banerjee, Amiya K.

doi:10.1128/jvi.75.4.1984-1989.2001

Cited by 50 publications

(55 citation statements)

References 25 publications

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“…Human lung epithelial A549 cells were maintained in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum, penicillin, streptomycin, and glutamine. The RSV titer was monitored by plaque assay analysis with CV-1 cells as described earlier (7,14,15). Vaccinia virus (WR strain) was propagated as described earlier (17).…”

Section: Methodsmentioning

confidence: 99%

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Role of Human β-Defensin-2 during Tumor Necrosis Factor-α/NF-κB-mediated Innate Antiviral Response against Human Respiratory Syncytial Virus

Kota

Sabbah

Chang

et al. 2008

Journal of Biological Chemistry

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143

137

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Human respiratory syncytial virus (RSV) constitutes a highly pathogenic virus that infects lung epithelial cells to cause a wide spectrum of respiratory diseases. Our recent studies have revealed the existence of an interferon-␣/␤-independent, innate antiviral response against RSV that was dependent on activation of NF-B. We demonstrated that NF-B inducing pro-inflammatory cytokines like tumor necrosis factor-␣ (TNF) confers potent antiviral function against RSV in an NF-B-dependent fashion, independent of interferon-␣/␤. During our efforts to study this pathway, we identified HBD2 (human ␤-defensin-2), a soluble secreted cationic protein as an antiviral factor induced during NF-B-dependent innate antiviral activity in human lung epithelial cells. Our results demonstrated that HBD2 is induced by TNF and RSV in an NF-B-dependent manner. Induction of HBD2 in infected cells was mediated by the paracrine/autocrine action of TNF produced upon RSV infection. HBD2 plays a critical role during host defense, because purified HBD2 drastically inhibited RSV infection. We also show that the antiviral mechanism of HBD2 involves blocking of viral cellular entry possibly because of destabilization/disintegration of the viral envelope. The important role of HBD2 in the innate response was also evident from loss of antiviral activity of TNF upon HBD2 silencing by short interfering RNA. The in vivo physiological relevance of HBD2 in host defense was apparent from induction of murine ␤-defensin-4 (murine counterpart of HBD2) in lung tissues of RSV-infected mice. Thus, HBD2 functions as an antiviral molecule during NF-B-dependent innate antiviral immunity mediated by the autocrine/paracrine action of TNF.Nonsegmented negative strand RNA viruses (superfamily Mononegaviridae) constitute highly pathogenic human viruses that cause high morbidity. Human respiratory syncytial virus (RSV), 2 a nonsegmented negative strand virus, is an important human lung tropic respiratory tract pathogen causing high morbidity and mortality among infants, children, and elderly by manifesting disease states, including pneumonia, and bronchiolitis (1, 2). To date, no effective vaccine or antiviral therapy exists for RSV. Therefore, elucidation of innate immune antiviral response induced by RSV holds significant potential for development of effective antiviral therapies in the near future.The innate immune antiviral response against viruses represents an important host defense mechanism (3). Innate immunity includes the first line of defense by the host to combat virus infection before an orchestrated adaptive immune response involving immune cell priming, and antibody production is launched. Two key molecules regulating the innate antiviral function are interferon regulatory factors (IRFs) and NF-B (4). These two transcription factors are activated either individually or together in infected cells, resulting in the expression and production of interferon-␣/␤ (IFN), which are potent antiviral cytokines (5, 6). IFN produced from infected cells binds to their...

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Section: Methodsmentioning

confidence: 99%

“…Virus and Cells-RSV (A2 strain) and adenoviruses (expressing either GFP or IB-super-repressor) were propagated in CV-1 cells and 293 cells, respectively (7,14,15). RSV was purified by centrifugation on discontinuous sucrose gradients as described previously (16).…”

Section: Methodsmentioning

confidence: 99%

Role of Human β-Defensin-2 during Tumor Necrosis Factor-α/NF-κB-mediated Innate Antiviral Response against Human Respiratory Syncytial Virus

Kota

Sabbah

Chang

et al. 2008

Journal of Biological Chemistry

Self Cite

143

137

View full text Add to dashboard Cite

show abstract

“…These cells display properties of type II alveolar epithelial cells and also form polarized barriers when grown on permeable filters (10,(17)(18)(19). A549 cells were maintained in Ham's F-12K medium with 2 mM L-glutamine 1.5 g/L NaHCO 3 , 10% FBS, and 100 U of penicillin/streptomycin.…”

Section: Growth and Maintenance Of Epithelial Cellsmentioning

confidence: 99%

Polymorphonuclear Cell Transmigration Induced by Pseudomonas aeruginosa Requires the Eicosanoid Hepoxilin A3

Hurley

Siccardi

Mrsny

et al. 2004

The Journal of Immunology

161

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Lung inflammation resulting from bacterial infection of the respiratory mucosal surface in diseases such as cystic fibrosis and pneumonia contributes significantly to the pathology. A major consequence of the inflammatory response is the recruitment and accumulation of polymorphonuclear cells (PMNs) at the infection site. It is currently unclear what bacterial factors trigger this response and exactly how PMNs are directed across the epithelial barrier to the airway lumen. An in vitro model consisting of human PMNs and alveolar epithelial cells (A549) grown on inverted Transwell filters was used to determine whether bacteria are capable of inducing PMN migration across these epithelial barriers. A variety of lung pathogenic bacteria, including Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeruginosa are indeed capable of inducing PMN migration across A549 monolayers. This phenomenon is not mediated by LPS, but requires live bacteria infecting the apical surface. Bacterial interaction with the apical surface of A549 monolayers results in activation of epithelial responses, including the phosphorylation of ERK1/2 and secretion of the PMN chemokine IL-8. However, secretion of IL-8 in response to bacterial infection is neither necessary nor sufficient to mediate PMN transepithelial migration. Instead, PMN transepithelial migration is mediated by the eicosanoid hepoxilin A3, which is a PMN chemoattractant secreted by A549 cells in response to bacterial infection in a protein kinase C-dependent manner. These data suggest that bacterial-induced hepoxilin A3 secretion may represent a previously unrecognized inflammatory mechanism occurring within the lung epithelium during bacterial infections.

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“…Since HPIV3 infects the epithelial cells of lung as the primary target, we used A549 cells that are widely used as a model for airway epithelia (Bose et al, 2001;Gao et al, 2001). To test the antiviral activity of C5 and C7, A549 monolayer was infected with HPIV3 at an MOI of 0.3 for 48h in the presence of 0.05, 0.55 or 5.5μM of the two compounds.…”

Section: Inhibition Of Wild Type Hpiv3 Infection In Cultured Cellsmentioning

confidence: 99%

“…Genome synthesis from antigenome template occurs in a fashion similar to that of antigenome synthesis. The nucleocapsid of the virus is subsequently assembled in the cytoplasm and virus buds from the apical surface of the epithelial cells (Bose et al, 2001). …”

Section: Introductionmentioning

confidence: 99%

Inhibition of human parainfluenza virus type 3 infection by novel small molecules

Mao¹,

Thakur²,

Chattopadhyay³

et al. 2008

Antiviral Research

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Human parainfluenza virus type 3 (HPIV3) is an important respiratory tract pathogen of infants and children. There are no vaccines or antivirals currently approved for prevention or treatment of HPIV3 infection. Towards developing an antiviral therapy to combat HPIV3 infection, we have established a green fluorescent protein (GFP)-tagged HPIV3 infected-cell assay and used it for screening of a small molecule library obtained from ChemBridge Diver. Two novel small molecules (C5 and C7) which shared structural similarities were identified and their inhibitory effects on HPIV3 were confirmed in CV-1 and human lung epithelium A549 cells by plaque assay, Western blot and Northern blot analyses. C5 and C7 effectively prevented the cytopathic effect in cells infected with HPIV3, achieving IC 50 values of 2.36μM and 0.08μM, respectively, for infectious virus production. The inhibition appears to be at the primary transcriptional level of HPIV3 life cycle based on sequential time course test, binding and internalization assays, and finally by a minigenome transcription assay in cells as well as measuring viral transcripts in cells in the presence of anisomycin. Interestingly, vesicular stomatitis virus (VSV), another member of mononegavirales order, was also inhibited by these compounds, whereas poliovirus-a picornavirus was not. Use of these inhibitors has a strong potential to develop novel antiviral agents against this important human pathogen.

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Polarity of Human Parainfluenza Virus Type 3 Infection in Polarized Human Lung Epithelial A549 Cells: Role of Microfilament and Microtubule

Cited by 50 publications

References 25 publications

Role of Human β-Defensin-2 during Tumor Necrosis Factor-α/NF-κB-mediated Innate Antiviral Response against Human Respiratory Syncytial Virus

Role of Human β-Defensin-2 during Tumor Necrosis Factor-α/NF-κB-mediated Innate Antiviral Response against Human Respiratory Syncytial Virus

Polymorphonuclear Cell Transmigration Induced by Pseudomonas aeruginosa Requires the Eicosanoid Hepoxilin A3

Inhibition of human parainfluenza virus type 3 infection by novel small molecules

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