Ciliated Cultures From Patients With Primary Ciliary Dyskinesia Do Not Produce Nitric Oxide or Inducible Nitric Oxide Synthase During Early Infection

Smith, Claire M.; Fadaee-Shohada, Mina J.; Sawhney, Rounak; Baker, N. F.; Williams, Gwyneth; Hirst, Robert A.; Andrew, Peter W.; O’Callaghan, Christopher

doi:10.1378/chest.13-0159

Cited by 18 publications

(15 citation statements)

References 34 publications

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“…This disorder relates to dysfunction of motile cilia, which share much structural homology with primary cilia. In addition there is evidence that cells isolated from these patients have a reduced capacity to respond to infectious challenge [46]. Moreover, very recently, human disorders related to primary cilia dysfunction, including Jeune syndrome [47] and short rib polydactyly (SRP) syndrome type III [48] have been linked to WDR34, already linked to NFκB signalling [49].…”

Section: Discussionmentioning

confidence: 99%

The primary cilium influences interleukin-1β-induced NFκB signalling by regulating IKK activity

Wann

Chapple

Knight

2014

Cellular Signalling

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The primary cilium is an organelle acting as a master regulator of cellular signalling. We have previously shown that disruption of primary cilia assembly, through targeting intraflagellar transport, is associated with muted nitric oxide and prostaglandin responses to the inflammatory cytokine interleukin-1β (IL-1β). Here, we show that loss of the primary cilium disrupts specific molecular signalling events in cytosolic NFκB signalling. The induction of cyclooxygenase 2 (COX2) and inducible nitrous oxide synthase (iNOS) protein is abolished. Cells unable to assemble cilia exhibit unaffected activation of IκB kinase (IKK), but delayed and reduced degradation of IκB, due to diminished phosphorylation of inhibitor of kappa B (IκB) by IKK. This results in both delayed and reduced NFκB p65 nuclear translocation and nuclear transcript binding. We also demonstrate that heat shock protein 27 (hsp27), an established regulator of IKK, is localized to the ciliary axoneme and cellular levels are dramatically disrupted with loss of the primary cilium. These results suggest that the primary cilia compartment exerts influence over NFκB signalling. We propose that the cilium is a locality for regulation of the molecular events defining NFκB signalling events, tuning signalling as appropriate.

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Section: Discussionmentioning

confidence: 99%

The primary cilium influences interleukin-1β-induced NFκB signalling by regulating IKK activity

Wann

Chapple

Knight

2014

Cellular Signalling

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“…pneumoniae strain D39 (NCTC 7466) was chosen because it is a well-characterized strain. We have previously shown it to be virulent in in vivo (12) and ex vivo studies with ciliated epithelia (13,14). The D39 pneumolysin-deficient mutant (ΔPLY) and wild-type strains were propagated in tryptic soy broth medium as described previously (15).…”

Section: Virus and Bacterial Growth Conditionsmentioning

confidence: 99%

“…After 2 hours the supernatants were harvested and the bacteria recovered by centrifugation. Bacteria were resuspended in RNAlater (Sigma) and stored at 280 8 C. The concentration of airway inflammation markers in the airway surface liquid was measured as before (19)(20)(21) using a 96-well multispot assay (Meso Scale Discovery, MD) according to the manufacturer's instructions. Cytokines were measured using a human Th1/Th2 standard 10-spot plate and human chemokines were measured using a highband MS6000 10-spot plate, using SECTOR Imager 6000 (MSD, MD).…”

Section: Infection Of Airway Epithelial Cell Culturesmentioning

confidence: 99%

Respiratory Syncytial Virus Increases the Virulence of Streptococcus pneumoniae by Binding to Penicillin Binding Protein 1a. A New Paradigm in Respiratory Infection

Smith

Sandrini

Datta

et al. 2014

Am J Respir Crit Care Med

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128

140

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Rationale: Respiratory syncytial virus (RSV) and Streptococcus pneumoniae are major respiratory pathogens. Coinfection with RSV and S. pneumoniae is associated with severe and often fatal pneumonia but the molecular basis for this remains unclear.Objectives: To determine if interaction between RSV and pneumococci enhances pneumococcal virulence.Methods: We used confocal microscopy and Western blot to identify the receptors involved in direct binding of RSV and pneumococci, the effects of which were studied in both in vivo and in vitro models of infection. Human ciliated respiratory epithelial cell cultures were infected with RSV for 72 hours and then challenged with pneumococci. Pneumococci were collected after 2 hours exposure and changes in gene expression determined using quantitative real-time polymerase chain reaction.Measurements and Main Results: Following incubation with RSV or purified G protein, pneumococci demonstrated a significant increase in the inflammatory response and bacterial adherence to human ciliated epithelial cultures and markedly increased virulence in a pneumonia model in mice. This was associated with extensive changes in the pneumococcal transcriptome and significant up-regulation in the expression of key pneumococcal virulence genes, including the gene for the pneumococcal toxin, pneumolysin. We show that mechanistically this is caused by RSV G glycoprotein binding penicillin binding protein 1a.Conclusions: The direct interaction between a respiratory virus protein and the pneumococcus resulting in increased bacterial virulence and worsening disease outcome is a new paradigm in respiratory infection.Keywords: respiratory syncytial virus; pneumococcus; cilia; virulence; G protein Respiratory syncytial virus (RSV) and Streptococcus pneumoniae, also known as the pneumococcus, are major respiratory pathogens causing a huge global healthcare burden, predominantly in young children and the elderly (1). Global RSV disease burden is estimated at 64 million cases and 160,000 deaths every year (2). Pneumococcal pneumonia results in more than 1 million deaths each year worldwide with approximately half a million in children younger than 5 years (3). There is increasing evidence that RSV and the pneumococcus interact to increase the severity of respiratory disease (4-6). Seasonal increases in infections This article has an online supplement, which is accessible from this issue's table of contents at www.atsjournals.org This article has embedded videos, which play in place from both the online PDF or HTML versions. If you cannot view Flash videos on your device, please try playing the videos in the online PDF, or access the original uncompressed videos at http://www.atsjournals.org/doi/suppl/10.1164/rccm.201311-2110OC. To play, mouse over the image and click on the arrow that will appear in the center or on the lower left.

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“…A simpler tool that has proven of great value in the correct identification of affected individuals constitutes the measurement of nasal NO production. Ciliary function is known to be regulated by NO and epithelial cells produce NO through epithelial NO synthases (NOS) (79). In contrast to healthy controls, epithelial cells from PCD patients are known to be defective in their ability to induce NOS2 expression and produce NO under infectious challenge (79).…”

Section: Nasal Nitric Oxide and Primary Ciliary Dyskinesiamentioning

confidence: 99%

“…Ciliary function is known to be regulated by NO and epithelial cells produce NO through epithelial NO synthases (NOS) (79). In contrast to healthy controls, epithelial cells from PCD patients are known to be defective in their ability to induce NOS2 expression and produce NO under infectious challenge (79). In the nasal cavity large concentrations range of NO close to the ppm can be detected, with it predominantly arising primarily from the sinuses (80).…”

Section: Nasal Nitric Oxide and Primary Ciliary Dyskinesiamentioning

confidence: 99%

The evolving spectrum of ciliopathies and respiratory disease

Milla

2016

Current Opinion in Pediatrics

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Purpose of review Research on the biology of cilia, complex hair-like cellular organelles, has greatly informed our understanding of its crucial role in respiratory health and the pathogenesis of Primary Ciliary Dyskinesia (PCD), including the genetics behind this condition. This review will summarize the current state of the art in the field highlighting its clinical implications. Recent findings The genetics of PCD have exploded over the past few years as knowledge acquired from model systems has permitted the identification of genes that are key components of the ciliary apparatus and its function. In addition clinical criteria and diagnostic tools have emerged that are permitting more clear identification of affected individuals. Summary The rate of progress in the field continues to accelerate through international collaborative efforts and standardization of methods. Although the genetics behind PCD are complex given the large number of genes associated with disease as well as the large number of possible mutations even at the individual gene level, this knowledge is rapidly translating in improved diagnostics and hopefully in the near future in the identification of potential therapeutics.

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Ciliated Cultures From Patients With Primary Ciliary Dyskinesia Do Not Produce Nitric Oxide or Inducible Nitric Oxide Synthase During Early Infection

Cited by 18 publications

References 34 publications

The primary cilium influences interleukin-1β-induced NFκB signalling by regulating IKK activity

The primary cilium influences interleukin-1β-induced NFκB signalling by regulating IKK activity

Respiratory Syncytial Virus Increases the Virulence of Streptococcus pneumoniae by Binding to Penicillin Binding Protein 1a. A New Paradigm in Respiratory Infection

The evolving spectrum of ciliopathies and respiratory disease

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