MAPK and heat shock protein 27 activation are associated with respiratory syncytial virus induction of human bronchial epithelial monolayer disruption

Singh, Divyendu; McCann, Kelly L.; Imani, Farhad

doi:10.1152/ajplung.00097.2007

Cited by 63 publications

(59 citation statements)

References 46 publications

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“…Here, we show that RSV, a common respiratory virus associated with substantial morbidity, induces epithelial barrier dysfunction in a PKD-dependent manner. RSV infection causes barrier dysfunction without altering the expression of key junctional components and instead appears to induce disassembly of epithelial junction complexes by destabilizing the cortical actin cytoskeleton in a cor- infection reduced transepithelial electrical resistance across airway epithelial monolayers (25). Our results clearly show that RSV infection leads to marked disassembly of AJC, resulting in increased paracellular permeability ( Fig.…”

Section: Discussionmentioning

confidence: 52%

Sustained Protein Kinase D Activation Mediates Respiratory Syncytial Virus-Induced Airway Barrier Disruption

Rezaee

DeSando

Ivanov

et al. 2013

J Virol

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Understanding the regulation of airway epithelial barrier function is a new frontier in asthma and respiratory viral infections. Despite recent progress, little is known about how respiratory syncytial virus (RSV) acts at mucosal sites, and very little is known about its ability to influence airway epithelial barrier function. Here, we studied the effect of RSV infection on the airway epithelial barrier using model epithelia. 16HBE14o-bronchial epithelial cells were grown on Transwell inserts and infected with RSV strain A2. We analyzed (i) epithelial apical junction complex (AJC) function, measuring transepithelial electrical resistance (TEER) and permeability to fluorescein isothiocyanate (FITC)-conjugated dextran, and (ii) AJC structure using immunofluorescent staining. Cells were pretreated or not with protein kinase D (PKD) inhibitors. UV-irradiated RSV served as a negative control. RSV infection led to a significant reduction in TEER and increase in permeability. Additionally it caused disruption of the AJC and remodeling of the apical actin cytoskeleton. Pretreatment with two structurally unrelated PKD inhibitors markedly attenuated RSV-induced effects. RSV induced phosphorylation of the actin binding protein cortactin in a PKD-dependent manner. UV-inactivated RSV had no effect on AJC function or structure. Our results suggest that RSV-induced airway epithelial barrier disruption involves PKD-dependent actin cytoskeletal remodeling, possibly dependent on cortactin activation. Defining the mechanisms by which RSV disrupts epithelial structure and function should enhance our understanding of the association between respiratory viral infections, airway inflammation, and allergen sensitization. Impaired barrier function may open a potential new therapeutic target for RSV-mediated lung diseases. R espiratory syncytial virus (RSV) is the most common respiratory pathogen in infants and young children (1) and an important cause of death in childhood (2). RSV has been identified as a source of morbidity and mortality in elderly and high-risk adults (3). RSV infects airway epithelial cells and is thought to cause tissue pathology by inducing the expression of proinflammatory mediators, leading to airway inflammation and, ultimately, an antiviral immune response (4). RSV also induces the expression of antiapoptotic genes and promotes epithelial cell survival, which is probably a strategy to ensure viral replication in infected cells (5).Emerging evidence points to a role for airway barrier dysfunction during respiratory viral infections (6), as well as in stable asthmatics (7). The airway barrier is made up of the surface mucus layer, as well as apical junction complexes (AJC) that regulate paracellular permeability (8). Previously we demonstrated that polyinosinic-polycytidylic acid [poly(I-C)], a synthetic doublestranded RNA and viral mimetic, induces potent breakdown of the airway epithelial AJC in a protein kinase D (PKD)-dependent manner (9). PKD, formerly known as PKC, is a serine/threonine protein kinase fami...

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Section: Discussionmentioning

confidence: 52%

Sustained Protein Kinase D Activation Mediates Respiratory Syncytial Virus-Induced Airway Barrier Disruption

Rezaee

DeSando

Ivanov

et al. 2013

J Virol

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“…Increased cell retraction was followed by increased H. somni invasion, since treatment of cells with BRSV prior to the addition of H. somni significantly increased paracellular migration of H. somni between BAT2 cells in a monolayer. BRSV may increase transmigration by increasing IbpA uptake by BAT2 cells, as suggested above, or BRSV may have a direct effect on the alveolar cell cytoskeleton, since rearrangement of actin filaments in human RSV-infected epithelial cells increases paracellular permeability (18,19). Thus, H. somni IbpA-and BRSV-induced cell retraction may partially explain the increased disease severity for animals dually infected with BRSV and H. somni (1).…”

Section: Discussionmentioning

confidence: 87%

Bovine Respiratory Syncytial Virus and Histophilus somni Interaction at the Alveolar Barrier

et al. 2013

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cOur previous studies showed that Histophilus somni and bovine respiratory syncytial virus (BRSV) act synergistically in vivo to cause more severe bovine respiratory disease than either agent alone causes. Since H. somni surface and secreted immunoglobulin binding protein A (IbpA) causes retraction of bovine alveolar type 2 (BAT2) cells and invasion between BAT2 cells in vitro, we investigated mechanisms of BRSV-plus-H. somni infection at the alveolar barrier. BRSV treatment of BAT2 cells prior to treatment with IbpA-rich H. somni concentrated culture supernatant (CCS) resulted in increased BAT2 cell rounding and retraction compared to those with either treatment alone. This mimicked the increased alveolar cell thickening in calves experimentally infected with BRSV followed by H. somni compared to that in calves infected with BRSV or H. somni alone. BRSVplus-H. somni CCS treatment of BAT2 cells also enhanced paracellular migration. The effect of matrix metalloproteinases (MMPs) was investigated as well because microarray analysis revealed that treatment with BRSV plus H. somni synergistically upregulated BAT2 cell expression of mmp1 and mmp3 compared to that in cells treated with either agent alone. Enzyme-linked immunosorbent assay (ELISA) confirmed that MMP1 and MMP3 protein levels were similarly upregulated. In collagen I and collagen IV (targets for MMP1 and MMP3, respectively) substrate zymography, digestion was increased with supernatants from dually treated BAT2 cells compared with those from singly treated cells. Enhanced breakdown of collagen IV in the basal lamina and of fibrillar collagen I in the adjacent interstitium in the dual infection may facilitate dissemination of H. somni infection. R espiratory infections are characteristically polymicrobial. We previously investigated the mechanisms of viral and bacterial synergy in bovine respiratory disease (BRD) (1) by infecting calves with bovine respiratory syncytial virus (BRSV) followed by infection with Histophilus somni or with either pathogen alone. The calves dually infected with BRSV and H. somni had the most severe disease and the highest serum IgE antibody responses to H. somni (1). Duration of pneumonia and persistence of H. somni in the lungs were also greatest in dually infected animals (1). Our earlier immunohistochemistry studies of experimental H. somni pneumonia showed that the bacteria are detected primarily in the alveoli at 24 h after intrabronchial inoculation (2). Since H. somni causes septicemia and its sequelae (3), it is likely that it crosses into the circulation over the alveolar barrier. We also showed that the toxic Fic (filamentation induced by cyclic AMP [cAMP]) motifs of the direct repeat domains (DR1 and DR2) of the immunoglobulin binding protein A (IbpA) cause bovine alveolar type 2 (BAT2) cells to retract in vitro, allowing paracellular migration (4). This cytotoxicity is due to adenylylation of host cell Rho GTPases by the Fic motif (5). Conservation of IbpA and Fic motifs in all tested H. somni disease isolates was c...

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“…Similar to IAPs, HSP27 also directly interacts with caspase 3 to inhibit cleavage (46) but has not been shown to directly possess antiviral activity. In fact, several viruses, including adenovirus (43), respiratory syncytial virus (36), and herpes simplex virus 1 (HSV-1) (28), target HSP27 activity to enhance viral replication directly through many of its nonapoptotic functions; however, further studies are needed to determine if HSP27 plays a direct role in promoting HCMV replication during myeloid differentiation. Nonetheless, our data demonstrate that HCMV targets the caspase 3 regulatory function of HSP27 during the early stages of infection to promote survival of the infected monocyte through the 48-h viability gate.…”

Section: Discussionmentioning

confidence: 99%

Human Cytomegalovirus Stimulates Monocyte-to-Macrophage Differentiation via the Temporal Regulation of Caspase 3

Chan

Nogalski

Yurochko

2012

J Virol

105

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Monocytes are primary targets for human cytomegalovirus (HCMV) infection and are proposed to be responsible for hematogenous dissemination of the virus. Biologically, monocytes have a short life span of 48 h in the circulation, a period of time during which monocytes must make a cell fate decision on whether to undergo apoptosis or differentiate into a macrophage. We have previously shown that HCMV infection stimulates monocyte-to-macrophage differentiation; however, the mechanism(s) by which HCMV-infected monocytes simultaneously navigate the 48-h “viability gate” and undergo macrophagic differentiation has remained elusive. Studies have demonstrated that the level of caspase 3 and 8 activities in monocytes may mediate the delicate balance between apoptosis and macrophage colony-stimulating factor (M-CSF)-induced myeloid differentiation. Here, we show that HCMV infection, unlike M-CSF treatment, does not induce caspase 8 activity to promote myeloid differentiation. However, HCMV infection does induce a temporal activation of caspase 3, with only a low level of active caspase 3 being observed after the 48-h viability checkpoint. Consistent with the role of a time-dependent activation of caspase 3 in promoting myeloid differentiation, the inhibition of caspase 3 blocked HCMV-induced monocyte-to-macrophage differentiation. Temporal transcriptome and functional analyses identified heat shock protein 27 (HSP27) and Mcl-1, two known regulators of caspase 3 activation, as being upregulated prior to the 48-h viability gate following HCMV infection. Using small interfering RNAs (siRNAs), we demonstrate that HCMV targets the rapid induction of HSP27 and Mcl-1, which cooperatively function to precisely control caspase 3 activity in order to allow for HCMV-infected monocytes to successfully traverse the 48-h cell fate decision checkpoint and commence macrophage maturation. Overall, this study highlights a unique regulatory mechanism employed by HCMV to tightly modulate the caspase 3 activity needed to promote myeloid differentiation, a key process in the viral dissemination and persistence strategy.

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MAPK and heat shock protein 27 activation are associated with respiratory syncytial virus induction of human bronchial epithelial monolayer disruption

Cited by 63 publications

References 46 publications

Sustained Protein Kinase D Activation Mediates Respiratory Syncytial Virus-Induced Airway Barrier Disruption

Sustained Protein Kinase D Activation Mediates Respiratory Syncytial Virus-Induced Airway Barrier Disruption

Bovine Respiratory Syncytial Virus and Histophilus somni Interaction at the Alveolar Barrier

Human Cytomegalovirus Stimulates Monocyte-to-Macrophage Differentiation via the Temporal Regulation of Caspase 3

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