Infection of cultured human tracheal epithelial cells by human parainfluenza virus types 2 and 3 (original) (raw)
Related papers
Journal of Virology, 2005
We constructed a human recombinant parainfluenza virus type 3 (rPIV3) that expresses enhanced green fluorescent protein (GFP) and used this virus, rgPIV3, to characterize PIV3 infection of an established in vitro model of human pseudostratified mucociliary airway epithelium (HAE). The apical surface of HAE was highly susceptible to rgPIV3 infection, whereas only occasional cells were infected when virus was applied to the basolateral surface. Infection involved exclusively ciliated epithelial cells. There was little evidence of virus-mediated cytopathology and no spread of the virus beyond the ciliated cell types. Infection of ciliated cells by rgPIV3 was sensitive to a neuraminidase specific for ␣2-6-linked sialic acid residues, but not to a neuraminidase that cleaves ␣2-3and ␣2-8-linked sialic acid residues. This provided evidence that rgPIV3 utilizes ␣2-6-linked sialic acid residues for initiating infection, a specificity also described for human influenza viruses. The PIV3 fusion (F) glycoprotein was trafficked exclusively to the apical surface of ciliated cells, which also was the site of release of progeny virus. F glycoprotein localized predominately to the membranes of the cilial shafts, suggesting that progeny viruses may bud from cilia per se. The polarized trafficking of F glycoprotein to the apical surface also likely restricts its interaction with neighboring cells and could account for the observed lack of cell-cell fusion. HAE derived from cystic fibrosis patients was not more susceptible to rgPIV3 infection but did exhibit limited spread of virus due to impaired movement of lumenal secretions due to compromised function of the cilia.
The Role of Human Parainfluenza Virus Infections in the Immunopathology of the Respiratory Tract
Current allergy and asthma reports, 2017
Viral infections are leading causes of both upper and lower airway acute illness in all age groups of healthy persons, and have also been implicated in the acute exacerbations of chronic respiratory disorders like asthma and COPD. Human rhinovirus, respiratory syncytial virus, influenza virus and coronavirus have been considered as the most important respiratory pathogens and relatively little attention has been paid to the role of parainfluenza viruses (hPIVs). Human parainfluenza viruses are single-stranded RNA viruses belonging to the paramyxovirus family that may evoke lower respiratory infections in infants, children and immunocompromised individuals. Among non-immune compromised adults, hPIV infection typically causes mild disease manifested as upper respiratory tract symptoms and is infrequently associated with severe croup or pneumonia. Moreover, hPIV infection may be associated with viral exacerbations of chronic airway diseases, asthma or COPD or chronic rhinosinusitis. In...
Journal of Inflammation, 2015
Background: Human parainfluenza virus type 3 (HPIV3), while infecting lower airway epithelial cells induces pneumonia and bronchiolitis in infants and children, and may lead to asthma exacerbations in children and adults. Respiratory viruses invading the airway epithelium activate innate immune response and induce inflammatory cytokine release contributing to the pathophysiology of upper and lower airway disorders. However, the effects of HPIV3 infection on nasal epithelial cells have not been well defined. The aim of this study was to evaluate the effect of the HPIV3 infection on cultured human nasal epithelial cells (HNECs) and the release of interferon gamma and other cytokines. Methods: RPMI 2650, a human nasal epithelial cell line was cultured into confluence and was infected with HPIV3 (MOI of 0.1, 0.01 and 0.001). The protein release into supernatants and mRNA expression of selected cytokines were assessed 24, 48 and 72 h after infection. Cytokine concentrations in supernatants were measured by ELISA and expression of cytokine mRNA in RPMI 2650 cells confirmed by real time RT-PCR analysis. Results: HNECs infection with HPIV3 did not induce cytotoxicity for at least 48 hours, but significantly increased IFN-γ protein concentration in the cell supernatants at 24 h and 48 h post infection (by 387% and 485% respectively as compared to mock infected cells). At 24 h a significant increase in expression of mRNA for IFNγ was observed. RANTES protein concentration and mRNA expression were significantly increased at 72 h after infection (mean protein concentration: 3.5 ± 1.4 pg/mL for 0.001 MOI, 10.8 ± 4.6 pg/mL for 0.01 MOI and 61.5 ± 18.4 pg/mL for 0.1 MOI as compared to 2.4 ± 1.3 pg/mL for uninfected cells). No measurable concentrations of TNF-α, IL-10, TSLP, IL-8, GM-CSF or eotaxin, were detected in virus infected cells supernatants. Conclusions: HPIV3 effectively infects upper airway epithelial cells and the infection is associated with induction of IFN-γ and generation of RANTES.
Journal of Biology and Today's World, 2016
The economic losses of infectious laryngotracheitis virus (ILTV) are prevented using attenuated live vaccines. Differential diagnosis of ILTV strains is still a critical problem in controlling programs. In this study, the embryonated chicken liver cell (ECL) serves as a host model to study virulence characteristics of ILTV strains. The permissivity of ECL cells to ILTV infection was investigated by assessing susceptibility of the cells to vaccine strain and virulent strain infections, analyzing the impact of viral infection on cell viability, and determining the host cellular factor X (FX) and cyclophilin A (CypA) at three passages. To evaluate the lytic replication dynamics of ILTV in infected cells the collected suspension of last passage of each strain was inoculated onto the dropped chorio-allantoic membrane of specific pathogen free eggs then checked for observing characteristic lesions. The results indicated that ECL cells are highly susceptible to attenuated vaccine strain ILTV infection. Upon infection, the strain showed faster replication kinetics in cell culture and marked cytopathic effects. Virulent strain was able to enter ECL cells but no infectious virus was produced at 3rd passage. The establishment of latency state was not confirmed by reactivation assay. In contrast to vaccine strain, cellular FX was also traced following virulent strain infection. The difference expression pattern of FX in ILTV strains-infected cells is most closely with the presence of cytopathic effects in culture. The embryonated chicken lung cell system may potentiate the relevant tool for differential diagnosis of ILTV strains.
Clinical Aspects of Bronchial Reactivity and Cell–Virus Interaction
Elsevier eBooks, 2005
Preschool children Respiratory syncytial virus, parainfluenza viruses, rhinovirus, coronaviruses Adolescents Rhinoviruses, mycoplasma pneumoniae, parainfluenza viruses, respiratory syncytial viruses Adults Rhinoviruses, parainfluenza viruses, mycoplasma pneumoniae, chlamydia pneumoniae
The Journal of allergy and clinical immunology, 2017
Leading etiologies of acute illnesses, respiratory viruses typically cause self-limited diseases, though severe complications can occur in fragile patients. Rhinoviruses, respiratory enteroviruses, influenza virus, respiratory syncytial viruses and coronaviruses are highly prevalent respiratory pathogens, but due to the lack of reliable animal models, their differential pathogenesis remains poorly characterized. To compare infections by respiratory viruses isolated from clinical specimens using reconstituted human airway epithelia. Tissues were infected with rhinoviruses RV-A55, RV-A49, RV-B48, RV-C8 and RV-C15, respiratory enterovirus EV-D68, influenza virus H3N2, respiratory syncytial virus RSV-B and coronavirus HCoV-OC43. Replication kinetics, cell tropism, impact on tissue integrity and cytokine secretion were compared. Virus adaptation and tissue response were assessed through RNA-sequencing. Rhinoviruses, RSV-B and HCoV-OC43 infected ciliated cells and caused no major cell dea...
Journal of Virology, 2001
In the present study we investigated the interaction of HPIV-3 with the type II alveolar human lung polarized epithelial A549 cells. Although HPIV-3 entry and budding were bidirectional from both the apical and the basolateral domains, HPIV-3 exhibited preferential entry and release from the apical pole. While disruption of the cellular actin microfilament and microtubule by cytochalasin D and nocodazole, respectively, had no effect on virus entry, disruption of the microtubule but not the microfilament inhibited HPIV-3 release.
Replication of non-respiratory viruses in tracheal organ cultures
PubMed, 1983
Employing Hoorn's technique, tracheal explant cultures were set from adult hamsters, rabbits and human foetuses. To determine the susceptibility of these cultures they were infected with nine different mainly non-respiratory viruses. Assay of virus was carried out in susceptible cell lines. The results of these studies indicated that herpes simplex type I (HSV-1) and vaccinia viruses multiplied in these cultures and caused ciliostasis. Herpes simplex type 2 (HSV-2) although multiplying in all, produced ciliostasis only in human foetal tracheal cultures. However, Chandipura (CHP), West Nile (WN), sandfly fever (SF-N) and polio-1 viruses multiplied without ciliostasis. These cultures did not support multiplication of Japanese encephalitis (JE) and Chikungunya (CHIK) viruses. The studies indicated that mammalian tracheal cultures support replication of the non-respiratory viruses. The continued and undiminished multiplication of viruses over long periods without ciliostasis suggests a role for the trachea in the transmission of viral infections by aerosol.
Resistance of differentiated human airway epithelium to infection by rhinovirus
American Journal of Physiology-Lung Cellular and Molecular Physiology, 2004
Virtually all in vitro studies of the effects of rhinovirus on human airway epithelium have used cells grown under conditions known to produce low levels of differentiation. The relevance of the results to native epithelium is questionable. Here we grew primary cultures of human tracheal or nasal epithelium under three conditions. One condition produced pseudostratified, mucociliary cells virtually indistinguishable from native epithelium. The other two conditions produced undifferentiated squamous cells lacking cilia. Cells were infected for 6 h with rhinovirus-16. After a 24-h incubation period, we determined levels of viral RNA in the cells, numbers of infectious viral particles released in the mucosal medium, expression of a variety of epithelial cytokines and other proteins, release of IL-6 and IL-8, and transepithelial electrical resistance and voltage. After infection, levels of viral RNA in the poorly differentiated cells were 30 or 130 times those in the differentiated. Fur...