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Associate Professor Brian Oliver

Biography

My scientific training began at the National Heart and Lung Institute, UK, where I mastered the isolation and in-vitro culture of several types of human lung cells. I then had further training in both molecular biology (two publications: we discovered a new gene in humans, and examined epigenetic programming events during embryogenesis) (University of Leeds) and then virology at Prof Sebastian Johnston’s laboratory at Imperial College, UK before commencing my PhD at The University of Sydney (supervised by Prof Judith Black). 

I am now head of the Respiratory Molecular Pathogenesis group with laboratory facilities located at both UTS and the Woolcock Institute . The work from my group is recognised to be amongst the best in the world, evidenced by selection for presentation at symposia at both national and large international conferences as well as resulting in prestigious publications.

Professional

Service to the field / professional activities

I accepted an invitation to join the Thoracic Society of Australia and New Zealand NSW executive committee in 2007, and have now become branch treasurer. In this role my aim is to improve collaboration between physicians and scientists by integrating basic research programs into the clinical arena.

In 2013, I was the driving force behind the establishment of the TSANZ NSW annual scientific meeting, which for the first time has brought respiratory researchers from across NSW together.

I am a member of the TSANZ, the European Respiratory Society, the American Thoracic Society and the American Physiological Society.  

Associate member F1000

Editor BioMed Research International 2013 onwards

Image of Brian Oliver
Associate Professor, School of Life Sciences
Core Member, CHT - Centre for Health Technologies
B Sc (Human Biology), M Sc (Medicine), Ph D
Member, European Respiratory Society
Senior Member, The Thoracic Society of Australia and New Zealand
Member, American Physiological Society
Member, American Thoracic Society
 

Research Interests

Brian Oliver is a NH&MRC career development fellow, who leads a productive team of researchers investigating the pathophysiology of respiratory diseases, with a particular emphasis on understanding basic mechanisms leading to disease exacerbations and progression.

My research can be divided into two main areas:

Respiratory infectious diseases

I am interested in understanding why respiratory infections can lead to the development of lung diseases, and also why the same infections can cause an acute increase in symptoms of preexisiting lung diseases.

These are examples of my research in this area.

My team were the first to demonstrate that primary human lung cells from people with asthma have an increased inflammatory response to rhinovirus infection (Resp Res 2006).  This increased response is virus specific, and related to differential transcription factor recruitment to inflammatory gene promoters.  Since rhinovirus-induced inflammation correlates with both the occurrence and severity of asthma exacerbations this finding helps to explain why exacerbations occur.

We discovered important cellular mechanisms which increase the risk of post-viral secondary infections (Thorax 2008). Specifically we found that virus-infected alveolar macrophages have both a marked inability to release bacterial-induced proinflammatory cytokines and impaired bacterial phagocytosis.

In a local collaboration, we were the first group in the world to show that exhaled breath contains viable respiratory viruses (CID 2008 & JMV 2009); a finding that has now been confirmed by other groups. This important finding is beginning to change the paradigms surrounding respiratory viral transmission.

My group have recently developed an in-vitro model of rhinovirus-induced asthma exacerbations (AJRCMB 2010) which is providing new insight into why β2-agonists, commonly used asthma medications, may be ineffective during exacerbations. Using this model we have recently discovered the cause of  β2-adronergic desensitisation (PLoS One 2012).

We have been investigating the potential reasons why viral infections may promote the development of asthma.  In asthma the airways are fibrotic, but the reason for this is not known.  We hypothesised that fibrosis may occur as a consequence of virus infection. My group has recently published our findings that primary airway cells from people with asthma produce a different panel of rhinovirus-induced ECM proteins in comparison to cells from people without asthma. Kuo C, et al Oliver BG. Rhinovirus infection induces extracellular matrix protein deposition in asthmatic and non-asthmatic airway smooth muscle cells A J Physiology – LCMP (2011) 300 L951-L957 

Respiratory Pathogenesis

My undergraduate degree in Human Biology sparked an interest which has now become one of my research foci.  I am intrigued why respiratory diseases develop, and why therapeutics often are not as effective as we would hope.

The following are examples of my research in this area

We have been investigating the β2-adronergic pathway, and its regulation, in airway smooth muscle cells from people with asthma, compared to people without asthma. we discovered that a key enzyme which regulates this pathway (PDE4) is overexpressed in asthma, functionally impairing the β2-adronergic pathway (Trian T, ….. and Oliver BG. PLoS ONE 6(5): e20000 2011 doi:10.1371/journal.pone.0020000).  I have also discovered that, contrary to observations in other species, PDE4 is not upregulated by β2-agonists, and therefore is not involved in β2-agonist-induced β2- adronergic tachiphylaxis (Niimi K, ….and Oliver BG). Am J Physiol Lung Cell Mol Physiol 302:L334-L342 2012).

We found that cigarette smoke, the major cause of COPD in developed countries, is a profibrotic stimulus to fibroblasts from only people with COPD.  The reason why fibroblasts only from people with COPD respond is intriguing and raises many questions regarding the pathogenesis of COPD.1. Krimmer DI, et al, Oliver BG. Matrix proteins from smoke exposed fibroblasts are proproliferative Am J Respir Cell (2012) 46 1 34-39. 

We have been investigating the aeitology of fibrosis in asthma, and have discovered that tumstatin, an endogenous regulator of angiogenesis, is missing from the airways of people with asthma. We tested the affect of addition of tumstatin using a chronic animal model of asthma, and found that airways hyperresponsiveness, inflammation and angiogenesis were inhibited - demonstrating the complex interaction between the matrix and lung pathophysiology.  Burgess, J. K.; el al; Oliver, BG. Reduction of tumstatin in asthmatic airways contributes to angiogenesis, inflammation, and hyperresponsiveness Am J Respir Crit Care Med (2010) 

There is increasing evidence that inflammation and fibrosis develop concurrently, but along independent pathways in lung diseases such as asthma and pulmonary fibrosis. To explore this in-vitro we examined the effect of simulating primary human lung epithelial, smooth muscle and fibroblasts with the pro-inflammatory leukotrienes and the anti-inflammatory prostaglandins.  We found the ECM deposition was induced by only prostaglandins supporting the notion that inflammation is not the driving force behind lung fibrosis. Van Ly D, et al, Oliver BG Prostaglandins but not leukotrienes alter extracellular matrix protein deposition & cytokine release in primary human airway smooth muscle cells and fibroblasts Am J Physiol Lung Cell Mol Physiol (2012) 303 L239-L250

My group was the first to show that primary human lung mesenchymal cells are capable of incorporating exogenously produced soluble extracellular matrix proteins into the deposited extracellular matrix around the cells.  This highlights the complexity of ECM deposition in-vivo, and the need to study multiple cell type in-vitro to understand how the ECM is formed in-vivo. Chen L, et al Oliver BG Differential regulation of extracellular matrix and soluble fibulin-1 levels by TGF-β1 in airway smooth muscle cells PLoS ONE (2013) 8:2

Publication metrics

In the last five years I have published 41 journal articles (60 career total), of which I was first/senior on 23 (with a major role in all publications), two book chapters and three patents. In the last 5 years I have had 4 publications in the top 2 highest ranked (mean IF 8.69) of 46 journals in the field respiratory system, and 2 in the top 2 highest ranked (mean IF 8.64) of 24 journals in the field allergy of the ISI Web of Knowledge 2011 JCR Science Edition. Using data form Google Scholar, my research papers have been cited 1367 times (the highest individual article has 207 citations), and when considering only papers published in the last five years my H index is 18, and my i10 index (the number of papers with at least 10 citations in the last 5 years) index is 30.

Can supervise: Yes
I supervise Honours, MSc and PhD students.  Please have a look at my research interests to get some idea of the types of projects which I offer.  
I teach in courses in the Faculty of Science

Chapters

Oliver, B.G., Burgess, J.K., Black, J. & Panettieri, R.A. 2013, 'Noncontractile Functions of Airway Smooth Muscle' in Middleton's Allergy: Principles and Practice: Eighth Edition, pp. 315-326.
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Black, J., Burgess, J., Oliver, B. & Moir, L. 2008, 'Altered Properties of Airway Smooth Muscle in Asthma' in Airway Smooth Muscle in Asthma and COPD: Biology and Pharmacology, pp. 181-199.
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Conferences

McAlinden, K., Chan, Y., Kota, A., Chen, H., Oliver, B. & Sharma, P. 2017, 'Maternal E-cigarette Vaping Enhances Development of Allergic Asthma In the Offspring', American Journal of Respiratory and Critical Care Medicine, American Thoracic Society.
Rationale: E-cigarettes (eCig) are being considered as an alternative to quit cigarette smoking (CS) while their long-term safety and effect on lung patho-physiology are not known. Maternal eCig-vaping may be considered as a safer CS-replacement during pregnancy. Thus the effect of maternal eCig vaping needs further assessment, particularly the effect this has on offspring and development of allergic asthma later in life. Combining mouse models of maternal vaping and allergic asthma and human airway smooth muscle cells (ASM) in vitro we tested whether maternal eCig vaping enhances features of allergic asthma in the offspring. Methods: Female BALB/c mice were vaped with either eCig vapour (± nicotine) or CS+eCig (+nicotine) or room air (control group). The eCig vaping was started prior to mating and continued during gestation and lactation while CS-exposure was used prior to mating and replaced with eCig during gestation and mating. The female offspring from these mothers were subjected to an ovalbumin (OVA)-induced allergic asthma model. 24 hours after the last aerosolized OVA or saline challenge, lung function measurements were performed using flexiVent (Scireq, Canada) to increasing concentration of methacholine (MCh). Airway inflammation was assessed by counting total immune cell influx in BAL fluid. Human ASM cells were treated with varying concentrations of eCig liquid condensate and key parameters of mitochondrial function were measured with a Seahorse XF analyzer. Results: Repeated allergen-exposure induced Th2-driven inflammation in OVA-exposed mice, characterized by massive influx of leukocytes predominantly eosinophils (OVA: 3x105±8.3x104 vs Saline: 1.1x102±1x102) and to some extent neutrophils (OVA: 1.3x104±4.4x103 vs Saline: 1.3x102±1.1x102) into the airways. The effect of allergen on airway eosinophilia was significantly enhanced in the offsprings from eCig OVA (+Nic)-exposed mothers when compared with eCig OVA (-Nic) or CS+eCig animals. OVA-exposed ...
Kota, A., Xenaki, D., Deshpande, D., Oliver, B. & Sharma, P. 2017, 'ASK1 Inhibition Prevented Mitogen-Induced Human Airway Smooth Muscle Growth in Chronic Obstructive Pulmonary Disease', American Journal of Respiratory and Critical Care Medicine, American Thoracic Society.
Mitchell, A.B., Glanville, A.R., Peters, M., Morgan, L.C. & Oliver, B.G. 2016, 'RESPIRATORY VIRUSES ARE COMMONLY DETECTED INASYMPTOMATIC ASTHMA AND COPD PATIENTS', Respirology, Wiley: 12 months, pp. 91-91.
Tovey, E.R., Liu-Brennan, D., Rimmer, J.S. & Oliver, B.G. 2014, 'New methods for measuring the time course of personal exposure to biological particles including aeroallergens', Indoor Air 2014 - 13th International Conference on Indoor Air Quality and Climate, pp. 926-931.
We present early data on two methods to monitor personal allergen exposure.

Journal articles

Faiz, A., Donovan, C., Nieuwenhuis, M.A.E., van den Berge, M., Postma, D.S., Yao, S., Park, C.Y., Hirsch, R., Fredberg, J.J., Tjin, G., Halayko, A.J., Rempel, K.L., Ward, J.P.T., Lee, T., Bossé, Y., Nickle, D.C., Obeidat, M., Vonk, J.M., Black, J.L., Oliver, B.G., Krishnan, R., McParland, B., Bourke, J.E. & Burgess, J.K. 2017, 'Latrophilin receptors: Novel bronchodilator targets in asthma', Thorax, vol. 72, pp. 74-82.
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© 2016 BMJ Publishing Group Ltd & British Thoracic Society.Background Asthma affects 300 million people worldwide. In asthma, the major cause of morbidity and mortality is acute airway narrowing, due to airway smooth muscle (ASM) hypercontraction, associated with airway remodelling. However, little is known about the transcriptional differences between healthy and asthmatic ASM cells. Objectives To investigate the transcriptional differences between asthmatic and healthy airway smooth muscle cells (ASMC) in culture and investigate the identified targets using in vitro and ex vivo techniques. Methods Human asthmatic and healthy ASMC grown in culture were run on Affymetrix_Hugene_1.0_ST microarrays. Identified candidates were confirmed by PCR, and immunohistochemistry. Functional analysis was conducted using in vitro ASMC proliferation, attachment and contraction assays and ex vivo contraction of mouse airways. Results We suggest a novel role for latrophilin (LPHN) receptors, finding increased expression on ASMC from asthmatics, compared with non-asthmatics in vivo and in vitro, suggesting a role in mediating airway function. A single nucleotide polymorphism in LPHN1 was associated with asthma and with increased LPHN1 expression in lung tissue. When activated, LPHNs regulated ASMC adhesion and proliferation in vitro, and promoted contraction of mouse airways and ASMC. Conclusions Given the need for novel inhibitors of airway remodelling and bronchodilators in asthma, the LPHN family may represent promising novel targets for future dual therapeutic intervention.
Patel, B.S., Rahman, M.M., Baehring, G., Xenaki, D., Tang, F.S.-.M., Oliver, B.G. & Ammit, A.J. 2017, 'Roflumilast N-Oxide in Combination with Formoterol Enhances the Antiinflammatory Effect of Dexamethasone in Airway Smooth Muscle Cells.', Am J Respir Cell Mol Biol, vol. 56, no. 4, pp. 532-538.
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Roflumilast is an orally active phosphodiesterase 4 inhibitor approved for use in chronic obstructive pulmonary disease. Roflumilast N-oxide (RNO) is the active metabolite of roflumilast and has a demonstrated antiinflammatory impact in vivo and in vitro. To date, the effect of RNO on the synthetic function of airway smooth muscle (ASM) cells is unknown. We address this herein and investigate the effect of RNO on 2-adrenoceptor-mediated, cAMP-dependent responses in ASM cells in vitro, and whether RNO enhances steroid-induced repression of inflammation. RNO (0.001-1,000 nM) alone had no effect on AMP production from ASM cells, and significant potentiation of the long-acting 2-agonist formoterol-induced cAMP could only be achieved at the highest concentration of RNO tested (1,000 nM). At this concentration, RNO exerted a small, but not significantly different, potentiation of formoterol-induced expression of antiinflammatory mitogen-activated protein kinase phosphatase 1. Consequently, tumor necrosis factor-induced IL-8 secretion was unaffected by RNO in combination with formoterol. However, because there was the potential for phosphodiesterase 4 inhibitors and long-acting 2-agonists to interact with corticosteroids to achieve superior antiinflammatory efficacy, we examined whether RNO, alone or in combination with formoterol, enhanced the antiinflammatory effect of dexamethasone by measuring the impact on IL-8 secretion. Although RNO alone did not significantly enhance the cytokine repression achieved with steroids, RNO in combination with formoterol significantly enhanced the antiinflammatory effect of dexamethasone in ASM cells. This was linked to increased mitogen-activated protein kinase phosphatase 1 expression in ASM cells, suggesting that a molecular mechanism is responsible for augmented antiinflammatory actions of combination therapeutic approaches that include RNO.
Chan, Y.L., Saad, S., Al-Odat, I., Oliver, B.G., Pollock, C., Jones, N.M. & Chen, H. 2017, 'Maternal L-Carnitine Supplementation Improves Brain Health in Offspring from Cigarette Smoke Exposed Mothers.', Front Mol Neurosci, vol. 10, p. 33.
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Maternal cigarette smoke exposure (SE) causes detrimental changes associated with the development of chronic neurological diseases in the offspring as a result of oxidative mitochondrial damage. Maternal L-Carnitine administration has been shown to reduce renal oxidative stress in SE offspring, but its effect in the brain is unknown. Here, we investigated the effects of maternal L-Carnitine supplementation on brain markers of oxidative stress, autophagy, mitophagy and mitochondrial energy producing oxidative phosphorylation (OXPHOS) complexes in SE offspring. Female Balb/c mice (8 weeks) were exposed to cigarette smoke prior to mating, during gestation and lactation with or without L-Carnitine supplementation (1.5 mM in drinking water). In 1 day old male SE offspring, brain mitochondrial damage was suggested by increased mitochondrial fusion and reduced autophagosome markers; whereas at 13 weeks, enhanced brain cell damage was suggested by reduced fission and autophagosome markers, as well as increased apoptosis and DNA fragmentation markers, which were partially reversed by maternal L-Carnitine supplementation. In female SE offspring, enhanced mitochondrial regeneration was suggested by decreased fission and increased fusion markers at day 1. At 13 weeks, there was an increase in brain energy demand, oxidative stress and mitochondrial turnover, reflected by the protein changes of OXPHOS complex, fission and autophagosome markers, as well as the endogenous antioxidant, which were also partially normalized by maternal L-Carnitine supplementation. However, markers of apoptosis and DNA fragmentation were not significantly changed. Thus L-Carnitine supplementation may benefit the brain health of the offspring from smoking mothers.
Sharma, P., Yi, R., Nayak, A., Wang, N., Knight, M.J., Pan, S., Oliver, B. & Deshpande, D.A. 2017, 'Bitter Taste Receptor Agonists Mitigate Features of Allergic Asthma in MiceRoslyn Yi, Ajay Nayak, Nadan Wang, Francesca Tang, Morgan J Knight, Shi Pan, Brian Oliver, and Deepak Deshpande', Scientific Reports.
Sharma, P., Kota, A., Deshpande, D., Haghi, M. & Oliver, B. 2017, 'Autophagy and airway fibrosis: Is there a link?', F1000 Research, vol. 6, no. 409.
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In the past decade, an emerging process named 'autophagy has generated intense interest in many chronic lung diseases. Tissue remodeling and fibrosis is a common feature of many airway diseases, and current therapies do not prevent or reverse these structural changes. Autophagy has evolved as a conserved process for bulk degradation and recycling of cytoplasmic components to maintain basal cellular homeostasis and healthy organelle populations in the cell. Furthermore, autophagy serves as a cell survival mechanism and can also be induced by chemical and physical stress to the cell. Accumulating evidence demonstrates that autophagy plays an essential role in vital cellular processes, including tissue remodeling. This review will discuss some of the recent advancements made in understanding the role of this fundamental process in airway fibrosis with emphasis on airway remodeling, and how autophagy can be exploited as a target for airway remodeling in asthma and chronic obstructive pulmonary disease.
Rumzhum, N.N., Rahman, M.M., Oliver, B.G. & Ammit, A.J. 2016, 'Effect of Sphingosine 1-Phosphate on Cyclo-Oxygenase-2 Expression, Prostaglandin E2 Secretion, and 2-Adrenergic Receptor Desensitization.', American journal of respiratory cell and molecular biology, vol. 54, no. 1, pp. 128-135.
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Tachyphylaxis of the 2-adrenergic receptor limits the efficacy of bronchodilatory 2-agonists in respiratory disease. Cellular studies in airway smooth muscle (ASM) have shown that inflammatory mediators and infectious stimuli reduce 2-adrenergic responsiveness in a cyclo-oxygenase (COX)-2-mediated, prostaglandin E2 (PGE2)-dependant manner. Herein, we show that sphingosine 1-phosphate (S1P), a bioactive sphingolipid that plays an important role in pathophysiology of asthma, also induces 2-adrenergic receptor desensitization in bronchial ASM cells and exerts hyporesponsiveness to 2-agonists. We treated ASM cells with S1P (1 M) for up to 24 hours and then examined the temporal kinetics of COX-2 mRNA expression, protein up-regulation, and PGE2 secretion. S1P significantly enhanced COX-2 expression and PGE2 secretion, and this was repressed by the selective COX-2 inhibitor celecoxib, the corticosteroid dexamethasone, or small interfering RNA (siRNA) knockdown of COX-2 expression. In combination with another proinflammatory mediator found elevated in asthmatic airways, the cytokine TNF-, we observed that S1P-induced COX-2 mRNA expression and protein up-regulation and PGE2 secretion from ASM cells were significantly enhanced. Notably, S1P induced heterologous 2-adrenergic desensitization, as measured by inhibition of cyclic adenosine monophosphate production in response to the short-acting 2-agonist, salbutamol, and the long-acting 2-agonist, formoterol. Taken together, these data indicate that S1P represses 2-adrenergic activity in ASM cells by increasing COX-2-mediated PGE2 production, and suggest that this bioactive sphingolipid found elevated in asthma may contribute to 2-adrenergic desensitization.
Stelzer-Braid, S., Tovey, E.R., Willenborg, C.M., Toelle, B.G., Ampon, R., Garden, F.L., Oliver, B.G., Strachan, R., Belessis, Y., Jaffe, A., Reddel, H.K., Crisafulli, D., Marks, G.B. & Rawlinson, W.D. 2016, 'Absence of back to school peaks in human rhinovirus detections and respiratory symptoms in a cohort of children with asthma', Journal of Medical Virology, vol. 88, no. 4, pp. 578-587.
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© 2016 Wiley Periodicals, Inc. Much of what is known about the seasonality of human rhinovirus (hRV) infections has been learned from the study of acute asthma exacerbations presenting to emergency care, including those among children at the start of the school term. Much less is known about the patterns of hRVs in the community. In this study, viruses and day-to-day symptoms of asthma and colds were monitored twice weekly in 67 children with asthma aged 5-12 years, over a 15 month period in Sydney, Australia. Overall hRV was detected in 314/1232 (25.5%) of nasal wash samples and 142/1231 (11.5%) of exhaled breath samples; of these, 231 and 24 respectively were genotyped. HRVs were detected with similar prevalence rate throughout the year, including no peak in hRV prevalence following return to school. No peaks were seen in asthma and cold symptoms using twice-weekly diary records. However, over the same period in the community, there were peaks in asthma emergency visits both at a large local hospital and in state-wide hospitalizations, following both return to school (February) and in late autumn (May) in children of the same age. This study suggests that hRV infections are common throughout the year among children, and differences in virus prevalence alone may not account for peaks in asthma symptoms.
Tang, F.S.M., Van Ly, D., Spann, K., Reading, P.C., Burgess, J.K., Hartl, D., Baines, K.J. & Oliver, B.G. 2016, 'Differential neutrophil activation in viral infections: Enhanced TLR-7/8-mediated CXCL8 release in asthma.', Respirology (Carlton, Vic.), vol. 21, no. 1, pp. 172-179.
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Respiratory viral infections are a major cause of asthma exacerbations. Neutrophils accumulate in the airways and the mechanisms that link neutrophilic inflammation, viral infections and exacerbations are unclear. This study aims to investigate anti-viral responses in neutrophils from patients with and without asthma and to investigate if neutrophils can be directly activated by respiratory viruses.Neutrophils from peripheral blood from asthmatic and non-asthmatic individuals were isolated and stimulated with lipopolysaccharide (LPS) (1g/mL), f-met-leu-phe (fMLP) (100nM), imiquimod (3g/mL), R848 (1.5g/mL), poly I:C (10g/mL), RV16 (multiplicity of infection (MOI)1), respiratory syncytial virus (RSV) (MOI1) or influenza virus (MOI1). Cell-free supernatants were collected after 1h of neutrophil elastase (NE) and matrix metalloproteinase (MMP)-9 release, or after 24h for CXCL8 release.LPS, fMLP, imiquimod and R848 stimulated the release of CXCL8, NE and MMP-9 whereas poly I:C selectively induced CXCL8 release only. R848-induced CXCL8 release was enhanced in neutrophils from asthmatics compared with non-asthmatic cells (P<0.01). RSV triggered the release of CXCL8 and NE from neutrophils, whereas RV16 or influenza had no effect.Neutrophils release CXCL8, NE and MMP-9 in response to viral surrogates with R848-induced CXCL8 release being specifically enhanced in asthmatic neutrophils. Toll-like receptor (TLR7/8) dysregulation may play a role in neutrophilic inflammation in viral-induced exacerbations.
Rumzhum, N.N., Patel, B.S., Prabhala, P., Gelissen, I.C., Oliver, B.G. & Ammit, A.J. 2016, 'IL-17A increases TNF--induced COX-2 protein stability and augments PGE2 secretion from airway smooth muscle cells: impact on 2 -adrenergic receptor desensitization.', Allergy, vol. 71, no. 3, pp. 387-396.
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IL-17A plays an important role in respiratory disease and is a known regulator of pulmonary inflammation and immunity. Recent studies have linked IL-17A with exacerbation in asthma and COPD. We have shown that the enzyme cyclooxygenase-2 (COX-2) and its prostanoid products, prostaglandin E2 (PGE2 ) in particular, are key contributors in in vitro models of infectious exacerbation; however, the impact of IL-17A was not known.We address this herein and show that IL-17A induces a robust and sustained upregulation of COX-2 protein and PGE2 secretion from airway smooth muscle (ASM) cells. COX-2 can be regulated at transcriptional, post-transcriptional and/or post-translational levels. We have elucidated the underlying molecular mechanisms responsible for the sustained upregulation of TNF--induced COX-2 by IL-17A in ASM cells and show that is not via increased COX-2 gene expression. Instead, TNF--induced COX-2 upregulation is subject to regulation by the proteasome, and IL-17A acts to increase TNF--induced COX-2 protein stability as confirmed by cycloheximide chase experiments. In this way, IL-17A acts to amplify the COX-2-mediated effects of TNF- and greatly enhances PGE2 secretion from ASM cells.As PGE2 is a multifunctional prostanoid with diverse roles in respiratory disease, our studies demonstrate a novel function for IL-17A in airway inflammation by showing for the first time that IL-17A impacts on the COX-2/PGE2 pathway, molecules known to contribute to disease exacerbation.
Patel, B.S., Rahman, M.M., Rumzhum, N.N., Oliver, B.G., Verrills, N.M. & Ammit, A.J. 2016, 'Theophylline Represses IL-8 Secretion From ASM Cells Independently of PDE Inhibition: Novel Role as a PP2A Activator.', American journal of respiratory cell and molecular biology, vol. 54, no. 6, pp. 792-801.
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Theophylline is an old drug experiencing a renaissance due to its beneficial anti-inflammatory effects in chronic respiratory diseases, such as asthma and COPD. Multiple modes of anti-inflammatory action have been reported, including inhibition of the enzymes that degrade cAMP - phosphodiesterase (PDE). Utilizing primary cultures of airway smooth muscle (ASM) cells, we recently revealed that PDE4 inhibitors can potentiate the anti-inflammatory action of 2-agonists by augmenting cAMP-dependent expression of the phosphatase that deactivates MAPK - MKP-1. Therefore the aim of this study was to address whether theophylline repressed cytokine production in a similar, PDE-dependent, MKP-1-mediated manner. Notably, theophylline did not potentiate cAMP release from ASM cells treated with the long-acting 2-agonist formoterol. Moreover, theophylline (0.1-10 &micro;M) did not increase formoterol-induced MKP-1 mRNA expression nor protein upregulation; consistent with the lack of cAMP generation. However, theophylline (at 10 &micro;M) was anti-inflammatory and repressed secretion of the neutrophil chemoattractant cytokine, IL-8, produced in response to tumor necrosis factor (TNF). Because theophylline's effects were independent of PDE4 inhibition or anti-inflammatory MKP-1, we then wished to elucidate novel mechanisms responsible. We investigated the impact of theophylline on protein phosphatase 2A (PP2A), a master controller of multiple inflammatory signaling pathways, and show that theophylline increases TNF-induced PP2A activity in ASM cells. Confirmatory results were obtained in A549 lung epithelial cells. PP2A activators have beneficial effects in ex vivo and in vivo models of respiratory disease. Thus, our study is the first to link theophylline with PP2A activation as a novel mechanism to control respiratory inflammation.
Oliver, B.G.G. & Black, J. 2016, 'Asthma: Airways that are hyperactive by design', American Journal of Respiratory and Critical Care Medicine, vol. 193, no. 6, pp. 596-598.
Chan, Y.L., Saad, S., Pollock, C., Oliver, B., Al-Odat, I., Zaky, A.A., Jones, N. & Chen, H. 2016, 'Impact of maternal cigarette smoke exposure on brain inflammation and oxidative stress in male mice offspring.', Scientific reports, vol. 6, p. 25881.
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Maternal cigarette smoke exposure (SE) during gestation can cause lifelong adverse effects in the offspring's brain. Several factors may contribute including inflammation, oxidative stress and hypoxia, whose changes in the developing brain are unknown. Female Balb/c mice were exposed to cigarette smoke prior to mating, during gestation and lactation. Male offspring were studied at postnatal day (P) 1, P20 and 13 weeks (W13). SE dams had reduced inflammatory mediators (IL-1, IL-6 and toll like receptor (TLR)4 mRNA), antioxidant (manganese superoxide dismutase (MnSOD)), and increased mitochondrial activities (OXPHOS-I, III and V) and protein damage marker nitrotyrosine. Brain hypoxia-inducible factor (HIF)1 and its upstream signalling molecule early growth response factor (EGR)1 were not changed in the SE dams. In the SE offspring, brain IL-1R, IL-6 and TLR4 mRNA were increased at W13. The translocase of outer mitochondrial membrane, and MnSOD were reduced at W13 with higher nitrotyrosine staining. HIF-1 was also increased at W13, although EGR1 was only reduced at P1. In conclusion, maternal SE increased markers of hypoxia and oxidative stress with mitochondrial dysfunction and cell damage in both dams and offspring, and upregulated inflammatory markers in offspring, which may render SE dams and their offspring vulnerable to additional brain insults.
Tovey, E.R., Liu-Brennan, D., Garden, F.L., Oliver, B.G., Perzanowski, M.S. & Marks, G.B. 2016, 'Time-Based Measurement of Personal Mite Allergen Bioaerosol Exposure over 24 Hour Periods.', PloS one, vol. 11, no. 5, p. e0153414.
Allergic diseases such as asthma and rhinitis are common in many countries. Globally the most common allergen associated with symptoms is produced by house dust mites. Although the bed has often been cited as the main site of exposure to mite allergens, surprisingly this has not yet been directly established by measurement due to a lack of suitable methods. Here we report on the development of novel methods to determine the pattern of personal exposure to mite allergen bioaerosols over 24-hour periods and applied this in a small field study using 10 normal adults. Air was sampled using a miniature time-based air-sampler of in-house design located close to the breathing zone of the participants, co-located with a miniature time-lapse camera. Airborne particles, drawn into the sampler at 2L/min via a narrow slot, were impacted onto the peripheral surface of a disk mounted on the hour-hand of either a 12 or 24 hour clock motor. The impaction surface was either an electret cloth, or an adhesive film; both novel for these purposes. Following a review of the time-lapse images, disks were post-hoc cut into subsamples corresponding to eight predetermined categories of indoor or outdoor location, extracted and analysed for mite allergen Der p 1 by an amplified ELISA. Allergen was detected in 57.2% of the total of 353 subsamples collected during 20 days of sampling. Exposure patterns varied over time. Higher concentrations of airborne mite allergen were typically measured in samples collected from domestic locations in the day and evening. Indoor domestic Der p 1 exposures accounted for 59.5% of total exposure, whereas total in-bed-asleep exposure, which varied 80 fold between individuals, accounted overall for 9.85% of total exposure, suggesting beds are not often the main site of exposure. This study establishes the feasibility of novel methods for determining the time-geography of personal exposure to many bioaerosols and identifies new areas for future technical develo...
Tang, F.S., Hansbro, P.M., Burgess, J.K., Ammit, A.J., Baines, K.J. & Oliver, B.G. 2016, 'A novel immunomodulatory function of neutrophils on rhinovirus-activated monocytes in vitro.', Thorax, vol. 71, no. 11, pp. 1039-1049.
Rhinovirus (RV) infections are the major precipitant of asthma exacerbations. While neutrophilic lung inflammation occurs during such infections, its role remains unclear. Neutrophilic inflammation is associated with increased asthma severity and steroid refractory disease. Neutrophils are vital for controlling infections but also have immunomodulatory functions. Previously, we found that neutrophils respond to viral mimetics but not replication competent RV. We aimed to investigate if neutrophils are activated and/or modulate immune responses of monocytes during RV16 infection.Primary human monocytes and autologous neutrophils were cocultured with or without RV16, in direct contact or separated by transwells. RV16-stimulated monocytes were also exposed to lysed neutrophils, neutrophil membrane components or soluble neutrophil intracellular components. Interleukin 6 (IL-6) and C-X-C motif (CXC)L8 mRNA and proteins were measured by quantitative PCR and ELISA at 24hours.RV16 induced IL-6 and CXCL8 in monocytes, but not neutrophils. RV16-induced IL-6 and CXCL8 from monocytes was reduced in the presence of live neutrophils. Transwell separation abolished the inhibitory effects. Lysed neutrophils inhibited RV16-induced IL-6 and CXCL8 from monocytes. Neutrophil intracellular components alone effectively inhibited RV16-induced monocyte-derived IL-6 and CXCL8. Neutrophil intracellular components reduced RV16-induced IL-6 and CXCL8 mRNA in monocytes.Cell contact between monocytes and neutrophils is required, and preformed neutrophil mediator(s) are likely to be involved in the suppression of cytokine mRNA and protein production. This study demonstrates a novel regulatory function of neutrophils on RV-activated monocytes in vitro, challenging the paradigm that neutrophils are predominantly proinflammatory.
Ing, M., Oliver, R.A., Oliver, B.G.G., Walsh, W.R. & Williamson, J.P. 2016, 'Evaluation of Transbronchial Lung Cryobiopsy Size and Freezing Time: A Prognostic Animal Study', Respiration, vol. 92, no. 1, pp. 34-39.
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&copy; 2016 S. Karger AG, BaselBackground: Transbronchial lung biopsy using a cryoprobe is a novel way of sampling lung parenchyma. Correlation of freezing time with biopsy size and complications has not been evaluated in vivo. Objectives: The primary aim of the study is to evaluate the correlation between transbronchial cryobiopsy freezing time and size. The secondary aims are to evaluate histological quality of the biopsy and evaluate procedure-associated complications. Methods: Transbronchial lung cryobiopsies were obtained from two anaesthetised sheep using a 1.9-mm cryoprobe inserted into a flexible bronchoscope under fluoroscopic guidance. Freezing times ranged from 1 to 6 s (n = 49). The cryobiopsies were evaluated histologically with respect to their size and quality. Complications of bleeding and pneumothorax were recorded. Results: The mean cross-sectional area of the cryobiopsy ranged from 4.7 &plusmn; 2.1 to 15.7 &plusmn; 15.3 mm2. There was a significant positive correlation between increasing freezing time and cryobiopsy cross-sectional area (p = 0.028). All biopsies contained lung tissue with preserved parenchyma. Crush and freeze artefacts were not observed and tissue architecture was intact in all specimens. Small blood vessels and terminal bronchioles were observed in 88% of specimens. All cryobiopsies caused nil or mild haemorrhage with the exception of only 1 episode of severe haemorrhage at 6 s freezing time. Pneumothoraces occurred at 2, 5 and 6 s freezing time and required chest tube insertion. The most significant haemorrhage and pneumothoraces occurred at 5 and 6 s. Our results suggest an initial freezing time of 3 s can provide the maximal biopsy size while minimising major complications. Conclusion: The optimal transbronchial cryobiopsy freezing time is initially 3 s. This time is associated with minimal complications and large artefact-free biopsies.
Chan, Y.L., Saad, S., Al-Odat, I., Zaky, A.A., Oliver, B., Pollock, C., Li, W., Jones, N.M. & Chen, H. 2016, 'Impact of maternal cigarette smoke exposure on brain and kidney health outcomes in female offspring.', Clinical and experimental pharmacology & physiology, vol. 43, pp. 1168-1176.
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Increased oxidative stress in the brain can lead to increased sympathetic tone that may further induce kidney dysfunction. Previously we have shown that maternal cigarette smoke exposure (SE) leads to significantly increased oxidative stress and inflammation in both brain and kidney, as well as reduced brain and kidney mitochondrial activity. This is closely associated with significant kidney underdevelopment and abnormal function in adulthood in the male offspring. This study aimed to investigate the impact of maternal SE on brain and kidney health in the female offspring. In this study, the mouse dams were exposed to 2 cigarettes, twice daily for 6 weeks prior to gestation, during pregnancy and lactation. Brains and kidneys from the female offspring were collected at 20 days (P20) and 13 weeks (W13) and were subject to further analysis. We found that mRNA expression of brain inflammatory markers interleukin-1 receptor and Toll-like receptor 4 were significantly increased in the SE offspring at both P20 and W13. Their brain mitochondrial activity markers were however increased at W13 with increased antioxidant activity. Kidney development and function in the female SE offspring were not different from the control offspring. We concluded that although brain inflammatory markers were upregulated in the SE female offspring, they were protected from some of the indicators of brain oxidative stress, such as endogenous antioxidant and mitochondrial dysfunction, as well as abnormal kidney development and function in adulthood. This article is protected by copyright. All rights reserved.
Chen, H., Chan, Y.L., Nguyen, L.T., Mao, Y., De Rosa, A., Beh, I.T., Chee, C., Oliver, B., Herok, G., Saad, S. & Gorrie, C. 2016, 'Moderate traumatic brain injury is linked to acute behaviour deficits and long term mitochondrial alterations.', Clinical and experimental pharmacology & physiology.
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Traumatic brain injury (TBI) remains one of the leading causes of death and disability worldwide. Mild TBI may lead to neuropsychiatric sequelae, including memory loss and motor impairment. Mitochondrial dysfunction and oxidative stress have a contributory role in several neurological disorders; however, their association with mitophagy in mild TBI is unclear. TBI was induced in female Sprague Dawley (SD) rats using a New York University Impactor (10g, impactor head 2.5mm diameter, weight drop 50mm) and compared to sham surgery controls. The novel object recognition and error ladder tests were performed at 24h and for 6 weeks post-injury, and the brains were examined histologically to confirm the extent of injury. Mitochondria Manganese Superoxide Dismutase (MnSOD) and the Oxidative Phosphorylation (OXPHOS) complexes I-V (CI-CV), as well as mitophagy markers, dynamin related protein 1 (DRP-1), LC3A/B and PTEN-induced putative kinase 1 (PINK-1), were measured in the penumbra by western blot. At 24 hours sham rats performed as expected on a novel object recognition test while TBI rats showed cognitive deficits at the early time points. TBI rats also showed more early motor deficits on a horizontal ladder, compared with the sham rats. MnSOD, OXPHOS CI, CIII and CV protein levels were significantly lower in the TBI group at 24 hours. DRP-1, LC3A/B I and II, and PINK-1 were increased at 6 weeks suggesting abnormal mitophagy. Moderate TBI caused immediate cognitive and mild motor functional deficits in the rats that did not persist. Reduced antioxidative capacity and possibly compromised mitochondrial function may affect the long-term functional recovery. This article is protected by copyright. All rights reserved.
Capistrano, S.J., Zakarya, R., Chen, H. & Oliver, B.G. 2016, 'Biomass Smoke Exposure Enhances Rhinovirus-Induced Inflammation in Primary Lung Fibroblasts.', International journal of molecular sciences, vol. 17, no. 9.
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Biomass smoke is one of the major air pollutants and contributors of household air pollution worldwide. More than 3 billion people use biomass fuels for cooking and heating, while other sources of exposure are from the occurrence of bushfires and occupational conditions. Persistent biomass smoke exposure has been associated with acute lower respiratory infection (ALRI) as a major environmental risk factor. Children under the age of five years are the most susceptible in developing severe ALRI, which accounts for 940,000 deaths globally. Around 90% of cases are attributed to viral infections, such as influenza, adenovirus, and rhinovirus. Although several epidemiological studies have generated substantial evidence of the association of biomass smoke and respiratory infections, the underlying mechanism is still unknown. Using an in vitro model, primary human lung fibroblasts were stimulated with biomass smoke extract (BME), specifically investigating hardwood and softwood types, and human rhinovirus-16 for 24 h. Production of pro-inflammatory mediators, such as IL-6 and IL-8, were measured via ELISA. Firstly, we found that hardwood and softwood smoke extract (1%) up-regulate IL-6 and IL-8 release (p 0.05). In addition, human rhinovirus-16 further increased biomass smoke-induced IL-8 in fibroblasts, in comparison to the two stimulatory agents alone. We also investigated the effect of biomass smoke on viral susceptibility by measuring viral load, and found no significant changes between BME exposed and non-exposed infected fibroblasts. Activated signaling pathways for IL-6 and IL-8 production by BME stimulation were examined using signaling pathway inhibitors. p38 MAPK inhibitor SB239063 significantly attenuated IL-6 and IL-8 release the most (p 0.05). This study demonstrated that biomass smoke can modulate rhinovirus-induced inflammation during infection, which can alter the severity of the disease. The mechanism by which biomass smoke exposure increases inflamm...
Mitchell, A.B., Mourad, B., Tovey, E., Buddle, L., Peters, M., Morgan, L. & Oliver, B.G. 2016, 'Spirometry filters can be used to detect exhaled respiratory viruses.', Journal of breath research, vol. 10, no. 4, p. 046002.
Respiratory viruses are very common in the community and contribute to the burden of illness for patients with chronic respiratory diseases, including acute exacerbations. Traditional sampling methods are invasive and problematic to repeat. Accordingly, we explored whether respiratory viruses could be isolated from disposable spirometry filters and whether detection of viruses in this context represented presence in the upper or lower respiratory tract. Discovery (n=53) and validation (n=49) cohorts were recruited from a hospital outpatient department during two different time periods. Spirometry mouthpiece filters were collected from all participants. Respiratory secretions were sampled from the upper and lower respiratory tract by nasal washing (NW), sputum, and bronchoalveolar lavage (BAL). All samples were examined using RT-PCR to identify a panel of respiratory viruses (rhinovirus, respiratory syncytial virus, influenza A, influenza B, parainfluenza virus 1, 2 & 3, and human metapneumovirus). Rhinovirus was quantified using qPCR. Paired filter-NW samples (n=29), filter-sputum samples (n=24), filter-BAL samples (n=39) and filter-NW-BAL samples (n=10) provided a range of comparisons. At least one virus was detected in any sample in 85% of participants in the discovery cohort versus 45% in the validation cohort. Overall, 72% of viruses identified in the paired comparator method matched those detected in spirometry filters. There was a high correlation between viruses identified in spirometry filters compared with viruses identified in both the upper and lower respiratory tract using traditional sampling methods. Our results suggest that examination of spirometry filters may be a novel and inexpensive sampling method for the presence of respiratory viruses in exhaled breath.
Ghadiri, M., Young, P.M., Jarolimek, W., Grau, G.E.R., Oliver, B.G.G. & Traini, D. 2016, 'The effect of non-specific tight junction modulators on the transepithelial transport of poorly permeable drugs across airway epithelial cells', Journal of Drug Targeting, pp. 1-8.
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&copy; 2016 Informa UK Limited, trading as Taylor & Francis GroupThe epithelial barrier in the respiratory system is a major obstacle for drug delivery to the systemic circulation in the lung. Epithelial barrier hinders the transport of large macromolecules or polar drugs. Essential components of this epithelial fence are physical intercellular structures termed tight junctions. Therefore, modulating tight junctions can enhance paracellular transport across epithelial barrier. In this study, the effect of some of non-specific tight junction modulators (TJMs); (Sodium (Na) decanoate, oleic acid and ethyleneglycol-bis-(-aminoethyl ether)-N, N-tetraacetic acid (EGTA)) with established effect on intestinal tight junctions was evaluated for its effects on bronchial epithelial cells (Calu-3 cells). It was demonstrated that the effect of TJMs especially Na decanoate resulted in a reversible opening of tight junctions evidenced by the decrease in the transepithelial resistance. It was also demonstrated that this reduction of TEER upon exposing the epithelial cells to the TJMs resulted in a significant increase in Flu-Na (paracellular marker) and PXS25 (anti-fibrotic compound) transepithelial transport through this barrier. In conclusion, among the investigated non-specific TJMs, Na decanoate fulfilled the requirements of an effective, non-toxic and reversible tight junction modulator for Calu-3 lung epithelial cells.
Mitchell, A.B., Oliver, B.G.G. & Glanville, A.R. 2016, 'Translational aspects of the human respiratory virome', American Journal of Respiratory and Critical Care Medicine, vol. 194, no. 12, pp. 1458-1464.
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&copy; 2016 by the American Thoracic Society.Despite the dominant role of community-acquired respiratory viruses as etiological agents of disease, there has been little focus to date on the translation of rapidly developing diagnostic modalities, such as next-generation sequencing techniques in the examination of lower respiratory tract samples. When applied, these techniques should inform strategies to both understand the nexus between health and disease states of the respiratory virome, and drive a paradigm shift in how the practicing pulmonologist views the conceptual framework of respiratory infections. The lower respiratory tract was once thought to be a sanctuary site from microbiological colonization owing to the efficacy of upper airway-protective mechanisms and the host mucosal barrier function of the lower airways, combined with both innate and adaptive immune responses.As a small number of recent studies confirm, this is a naive vision of the lung, the viral component of which parallels recent revelations from respiratory microbiome studies. Hence, it is now timely to revise our thinking regarding the constituents, diversity, and changing nature of the respiratory virome in health and disease. One area worthy of focus is the interface between community-acquired respiratory viruses and the respiratory virome to better understand the dynamics in acute infection, as well as the factors that may lead to viral persistence and chronic disease. Given recent advances in metagenomics, the tools are now at hand to accomplish these goals.
Liu, G., Cooley, M.A., Jarnicki, A.G., Hsu, A.C.-.Y., Nair, P.M., Haw, T.J., Fricker, M., Gellatly, S.L., Kim, R.Y., Inman, M.D., Tjin, G., Wark, P.A.B., Walker, M.M., Horvat, J.C., Oliver, B.G., Argraves, W.S., Knight, D.A., Burgess, J.K. & Hansbro, P.M. 2016, 'Fibulin-1 regulates the pathogenesis of tissue remodeling in respiratory diseases', JCI Insight, vol. 1, no. 9.
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Patel, B.S., Prabhala, P., Oliver, B.G. & Ammit, A.J. 2015, 'Inhibitors of Phosphodiesterase 4, but Not Phosphodiesterase 3, Increase 2-Agonist-Induced Expression of Antiinflammatory Mitogen-Activated Protein Kinase Phosphatase 1 in Airway Smooth Muscle Cells.', American journal of respiratory cell and molecular biology, vol. 52, no. 5, pp. 634-640.
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2-agonists are principally used in asthma to provide bronchodilation; however, they also have antiinflammatory properties, due, in part, to their ability to up-regulate mitogen-activated protein kinase phosphatase (MKP) 1 in a cAMP-dependent manner. Phosphodiesterases (PDEs) are attractive targets for potentiating the antiinflammatory response. There are 11 subfamilies of PDE enzymes; among these, inhibition of PDE3 and PDE4 are the main targets for airway smooth muscle (ASM). PDE enzymes are important intracellular regulators that catalyze the breakdown of cyclic adenosine monophosphate (cAMP) and/or 3',5'-cyclic guanosine monophosphate to their inactive forms. Given that MKP-1 is cAMP dependent, and inhibition of PDE acts to increase 2-agonist-induced cAMP, it is possible that the presence of PDE inhibitors may enhance 2-adrenoceptor-mediated responses. We address this herein by comparing the ability of a panel of inhibitors against PDE3 (cilostamide, cilostazol, milrinone) or PDE4 (cilomilast, piclamilast, rolipram) to increase cAMP, MKP-1 mRNA expression, and protein up-regulation in ASM cells induced in response to the 2-agonist formoterol. Our data show that inhibitors of PDE4, but not PDE3, increase 2-agonist-induced cAMP and induce MKP-1 mRNA expression and protein up-regulation. When cAMP was increased, there was a concomitant increase in MKP-1 levels and significant inhibition of TNF--induced CXCL8 (IL-8). This result was consistent with all PDE4 inhibitors examined but not for the PDE3 inhibitors. These findings reinforce cAMP-dependent control of MKP-1 expression, and suggest that PDE4 is the predominant PDE isoform responsible for formoterol-induced cAMP breakdown in ASM cells. Our study is the first to demonstrate that PDE4 inhibitors augment antiinflammatory effects of 2-agonists via increased MKP-1 expression in ASM cells.
Van Ly, D. & Oliver, B.G. 2015, 'Do we really need to keep redesigning 2-agonists for the management of asthma?', Current drug delivery, vol. 12, no. 1, pp. 9-15.
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There is an enormous drive to refine therapeutic designs and delivery systems, but in this review we ask if this is always the right direction? We choose to play devil's advocate, and argue that refining drug design is not always needed, and what is actually needed is a greater understanding of the biology of the disease. Here we focus on asthma and the 2-agonist group of bronchodilators as an example of how a class of therapeutic has been developed and continues to be developmentally refined. In this review, we define viral-induced exacerbations as the greatest cause of lung attacks and the most crucial time 2-agonist therapy is needed. We explore the reasons why 2-agonist therapy fails in patients with rhinovirus-induced exacerbations, and explain why further "engineered" 2-agonist therapies are likely to continue to fail in this subset of asthmatic population. We justify our perspective by returning to the biology that underlies the cause of disease and highlight the need for "more research" into alternative therapies for this population of asthmatic patients.
Tovey, E.R., Stelzer-Braid, S., Toelle, B.G., Oliver, B.G., Reddel, H.K., Willenborg, C.M., Belessis, Y., Garden, F.L., Jaffe, A., Strachan, R., Eyles, D., Rawlinson, W.D. & Marks, G.B. 2015, 'Rhinoviruses significantly affect day-to-day respiratory symptoms of children with asthma.', The Journal of allergy and clinical immunology, vol. 135, no. 3, pp. 663-9.e12.
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BACKGROUND: Viruses are frequently associated with acute exacerbations of asthma, but the extent to which they contribute to the level of day-to-day symptom control is less clear. OBJECTIVE: We sought to explore the relationship between viral infections, host and environmental factors, and respiratory symptoms in children. METHODS: Sixty-seven asthmatic children collected samples twice weekly for an average of 10 weeks. These included nasal wash fluid and exhaled breath for PCR-based detection of viral RNA, lung function measurements, and records of medication use and asthma and respiratory symptoms in the previous 3 days. Atopy, mite allergen exposure, and vitamin D levels were also measured. Mixed-model regression analyses were performed. RESULTS: Human rhinoviruses (hRVs) were detected in 25.5% of 1232 nasal samples and 11.5% of breath samples. Non-hRV viruses were detected in less than 3% of samples. hRV in nasal samples was associated with asthma symptoms (cough and phlegm: odds ratio = 2.0; 95% CI = 1.4-2.86, P = .0001; wheeze and chest tightness: odds ratio = 2.34, 95% CI = 1.55-3.52, P < .0001) and with cold symptoms, as reported concurrently with sampling and 3 to 4 days later. No differences were found between the 3 hRV genotypes (hRV-A, hRV-B, and hRV-C) in symptom risk. A history of inhaled corticosteroid use, but not atopic status, mite allergen exposure, or vitamin D levels, modified the association between viruses and asthma symptoms. CONCLUSION: The detection of nasal hRV was associated with a significantly increased risk of day-to-day asthma symptoms in children. Host, virus genotype, and environmental factors each had only a small or no effect on the relationship of viral infections to asthma symptoms.
Ge, Q., Chen, L., Jaffar, J., Argraves, W.S., Twal, W.O., Hansbro, P., Black, J.L., Burgess, J.K. & Oliver, B. 2015, 'Fibulin1C peptide induces cell attachment and extracellular matrix deposition in lung fibroblasts.', Scientific reports, vol. 5, p. 9496.
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Fibulin-1 is an extracellular matrix (ECM) protein, levels of which are elevated in serum and lung tissue from patients with idiopathic pulmonary fibrosis compared to healthy volunteers. Inhibition of fibulin-1C, one of four fibulin-1 isoforms, reduced proliferation and wound healing in human airway smooth muscle (ASM) cells. This study identified the bioactive region/s of fibulin-1C which promotes fibrosis. Seven fibulin-1C peptides were synthesized and used to pre-coat tissue culture plates before lung derived ASM cells and fibroblasts from patients with pulmonary fibrosis (PF), chronic obstructive pulmonary disease (COPD) or neither disease (Control) were plated. Peptide effects on in vitro measures of fibrosis: cell attachment, proliferation and viability, and ECM deposition, were examined. Among these peptides, peptide 1C1 (FBLN1C1) enhanced ASM cell and fibroblast attachment. FBLN1C1 increased mitochondrial activity and proliferation in fibroblasts. In addition, FBLN1C1 stimulated fibulin1 deposition in PF and COPD fibroblasts, and augmented fibronectin and perlecan deposition in all three groups. Peptides FBLN1C2 to FBLN1C7 had no activity. The active fibulin-1C peptide identified in this study describes a useful tool for future studies. Ongoing investigation of the role of fibulin-1 may reveal the mechanisms underlying the pathphysiology of chronic lung diseases.
Haghi, M., Traini, D., Wood, L.G., Oliver, B., Young, P.M. & Chrzanowski, W. 2015, 'A 'soft spot' for drug transport: modulation of cell stiffness using fatty acids and its impact on drug transport in lung model', JOURNAL OF MATERIALS CHEMISTRY B, vol. 3, no. 13, pp. 2583-2589.
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Haghi, M., Hittinger, M., Zeng, Q., Oliver, B.G., Traini, D., Young, P., Huwer, H., Schneider-Daum, N. & Lehr, C.M. 2015, 'Mono- and Cocultures of Bronchial and Alveolar Epithelial Cells Respond Differently to Proinflammatory Stimuli and Their Modulation by Salbutamol and Budesonide.', Mol Pharm, vol. 12, no. 8, pp. 2625-2632.
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The aim of this study was to investigate the changes in transport and effectiveness of salbutamol sulfate (SAL) and budesonide (BD) following stimulation with transforming growth factor- (TGF-) in mono- and coculture models of bronchial and alveolar epithelium. Primary bronchial and alveolar epithelial cells, grown at air interface on filters, either as monocultures or in coculture with airway smooth muscle cells or alveolar macrophages, respectively, were stimulated with TGF-. The biological response was modulated by depositing aerosolized SAL and BD on bronchial and alveolar models, respectively. Barrier integrity, permeability to fluorescein-Na, transport of the deposited drug, and the pharmacological response to SAL (cAMP and IL-8 levels) or BD (IL-6 and -8 levels) were measured. While stimulation with TGF- did not have any significant effect on the transepithelial electrical resistance and permeability to fluorescein-Na in mono- and coculture models, transport of SAL and BD were affected in cultures from some of the patients (6 out of 12 for bronchial and 2 out of 4 for alveolar cells). The bronchial coculture showed a better responsiveness to SAL in terms of cAMP release than the monoculture. In contrast, the difference between alveolar mono- and cocultures to TGF- mediated interleukin release and its modulation by BD was less pronounced. Our data point to intrinsic differences in the transport of, and responsiveness to, SAL and BD when epithelial cell cultures originate from different patients. Moreover, if the biological responses (e.g., IL-8, cAMP) involve communication between different cell types, coculture models are more relevant to measure such effects than monocultures.
Chan, Y.L., Saad, S., Simar, D., Oliver, B., McGrath, K., Reyk, D.V., Bertrand, P.P., Gorrie, C., Pollock, C. & Chen, H. 2015, 'Short term exendin-4 treatment reduces markers of metabolic disorders in female offspring of obese rat dams', International Journal of Developmental Neuroscience, vol. 46, pp. 67-75.
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Objectives Maternal obesity imposes significant health risks in the offspring including diabetes and dyslipidemia. We previously showed that the hypoglycaemic agent exendin-4 (Ex-4) administered from weaning can reverse the maternal impact of 'transmitted disorders' in such offspring. However daily injection for six-weeks was required and the beneficial effect may lapse upon drug withdrawal. This study aimed to investigate whether short term Ex-4 treatment during suckling period in a rodent model can reverse transmitted metabolic disorders due to maternal obesity. Methods Maternal obesity was induced in female Sprague Dawley rats by high-fat diet feeding for 6 weeks, throughout gestation and lactation. Female offspring were treated with Ex-4 (5 g/kg/day) between postnatal day (P) 4 and 14. Female offspring were harvested at weaning (P20). Lipid and glucose metabolic markers were measured in the liver and fat. Appetite regulators were measured in the plasma and hypothalamus. Results Maternal obesity significantly increased body weight, fat mass, and liver weight in the offspring. There was an associated inhibition of peroxisomal proliferator activated receptor gamma coactivator 1 (PGC1), increased fatty acid synthase (FASN) expression in the liver, and reduced adipocyte triglyceride lipase (ATGL) expression. It also increased the plasma gut hormone ghrelin and reduced glucagon-like peptide-1. Ex-4 treatment partially reversed the maternal impact on adiposity and impaired lipid metabolism in the offspring, with increased liver PGC1 and inhibition of FASN mRNA expression. Ex-4 treatment also increased the expression of a novel fat depletion gene a2-zinc-glycoprotein 1 in the fat tissue. Conclusion Short term Ex-4 treatment during the suckling period significantly improved the metabolic profile in the offspring from the obese mothers at weaning. Long-term studies are needed to follow such offspring to adulthood to examine the sustained effects of Ex-4 in p...
Herbert, C., Sebesfi, M., Zeng, Q.X., Oliver, B.G., Foster, P.S. & Kumar, R.K. 2015, 'Using multiple online databases to help identify microRNAs regulating the airway epithelial cell response to a virus-like stimulus.', Respirology (Carlton, Vic.), vol. 20, no. 8, pp. 1206-1212.
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Exacerbations of allergic asthma are often triggered by respiratory viral infections. We have previously shown that in a T-helper type 2 (Th2)-biased cytokine environment, mouse and human airway epithelial cells (AEC) exhibit increased expression of pro-inflammatory and anti-viral genes in response to synthetic double-stranded ribonucleic acid (dsRNA), a virus-like stimulus. This implies coordinated regulation of gene expression, suggesting possible involvement of microRNA. To investigate this, we developed a novel approach to identifying candidate microRNA using online databases, then confirmed their expression by quantitative real-time polymerase chain reaction (qRT-PCR).Using a list of genes of interest, defined on the basis of the previous study as being up-regulated in a Th2 environment, we searched mouse and human microRNA databases for possible regulatory microRNA, and selected 10 candidates that were conserved across species or predicted by more than one human database. Expression of these microRNA was tested by qRT-PCR, in primary human AEC pre-treated with Th2 cytokines and exposed to dsRNA.Expression of hsa-miR-139-5p, miR-423-5p and miR-542-3p was significantly decreased in Th2 pre-treated AEC, and miR-135a-5p exhibited a trend towards decreased expression. Further database searches confirmed that these microRNA regulated additional pro-inflammatory and anti-viral response genes for which expression had previously been shown to be up-regulated, confirming the validity of this approach.Our study demonstrates the value of using multiple online databases to identify candidate regulatory microRNA and provides the first evidence that in an allergic environment, microRNA may be important in altering the pro-inflammatory and anti-viral responses of human AEC during exacerbations of asthma.
Ng, H.Y., Oliver, B.G.G., Burgess, J.K., Krymskaya, V.P., Black, J.L. & Moir, L.M. 2015, 'Doxycycline reduces the migration of tuberous sclerosis complex-2 null cells - effects on RhoA-GTPase and focal adhesion kinase', Journal of Cellular and Molecular Medicine, vol. 19, no. 11, pp. 2633-2646.
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& Sons Ltd and Foundation for Cellular and Molecular Medicine. Lymphangioleiomyomatosis (LAM) is associated with dysfunction of the tuberous sclerosis complex (TSC) leading to enhanced cell proliferation and migration. This study aims to examine whether doxycycline, a tetracycline antibiotic, can inhibit the enhanced migration of TSC2-deficient cells, identify signalling pathways through which doxycycline works and to assess the effectiveness of combining doxycycline with rapamycin (mammalian target of rapamycin complex 1 inhibitor) in controlling cell migration, proliferation and wound closure. TSC2-positive and TSC2-negative mouse embryonic fibroblasts (MEF), 323-TSC2-positive and 323-TSC2-null MEF and Eker rat uterine leiomyoma (ELT3) cells were treated with doxycycline or rapamycin alone, or in combination. Migration, wound closure and proliferation were assessed using a transwell migration assay, time-lapse microscopy and manual cell counts respectively. RhoA-GTPase activity, phosphorylation of p70S6 kinase (p70S6K) and focal adhesion kinase (FAK) in TSC2-negative MEF treated with doxycycline were examined using ELISA and immunoblotting techniques. The enhanced migration of TSC2-null cells was reduced by doxycycline at concentrations as low as 20 pM, while the rate of wound closure was reduced at 2-59 M. Doxycycline decreased RhoA-GTPase activity and phosphorylation of FAK in these cells but had no effect on the phosphorylation of p70S6K, ERK1/2 or AKT. Combining doxycycline with rapamycin significantly reduced the rate of wound closure at lower concentrations than achieved with either drug alone. This study shows that doxycycline inhibits TSC2-null cell migration. Thus doxycycline has potential as an anti-migratory agent in the treatment of diseases with TSC2 dysfunction. &copy; 2015 John Wiley
Ge, Q., Zeng, Q., Tjin, G., Lau, E., Black, J.L., Oliver, B.G.G. & Burgess, J.K. 2015, 'Differential deposition of fibronectin by asthmatic bronchial epithelial cells', American Journal of Physiology - Lung Cellular and Molecular Physiology, vol. 309, no. 10, pp. L1093-L1102.
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&copy; 2015 the American Physiological Society. Altered ECM protein deposition is a feature in asthmatic airways. Fibronectin (Fn), an ECM protein produced by human bronchial epithelial cells (HBECs), is increased in asthmatic airways. This study investigated the regulation of Fn production in asthmatic or nonasthmatic HBECs and whether Fn modulated HBEC proliferation and inflammatory mediator secretion. The signaling pathways underlying transforming growth factor (TGF)-1-regulated Fn production were examined using specific inhibitors for ERK, JNK, p38 MAPK, phosphatidylinositol 3 kinase, and activin-like kinase 5 (ALK5). Asthmatic HBECs deposited higher levels of Fn in the ECM than nonasthmatic cells under basal conditions, whereas cells from the two groups had similar levels of Fn mRNA and soluble Fn. TGF-1 increased mRNA levels and ECM and soluble forms of Fn but decreased cell proliferation in both cells. The rate of increase in Fn mRNA was higher in nonasthmatic cells. However, the excessive amounts of ECM Fn deposited by asthmatic cells after TGF-1 stimulation persisted compared with nonasthmatic cells. Inhibition of ALK5 completely prevented TGF- 1-induced Fn deposition. Importantly, ECM Fn increased HBEC proliferation and IL-6 release, decreased PGE2 secretion, but had no effect on VEGF release. Soluble Fn had no effect on cell proliferation and inflammatory mediator release. Asthmatic HBECs are intrinsically primed to produce more ECM Fn, which when deposited into the ECM, is capable of driving remodeling and inflammation. The increased airway Fn may be one of the key driving factors in the persistence of asthma and represents a novel, therapeutic target.
Tang, F.S., Foxley, G.J., Gibson, P.G., Burgess, J.K., Baines, K.J. & Oliver, B.G. 2015, 'Altered Innate Immune Responses in Neutrophils from Patients with Well- and Suboptimally Controlled Asthma.', Mediators of inflammation, vol. 2015, p. 219374.
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Respiratory infections are a major cause of asthma exacerbations where neutrophilic inflammation dominates and is associated with steroid refractory asthma. Structural airway cells in asthma differ from nonasthmatics; however it is unknown if neutrophils differ. We investigated neutrophil immune responses in patients who have good (AGood) and suboptimal (ASubopt) asthma symptom control.Peripheral blood neutrophils from AGood (ACQ < 0.75, n = 11), ASubopt (ACQ > 0.75, n = 7), and healthy controls (HC) (n = 9) were stimulated with bacterial (LPS (1g/mL), fMLF (100nM)), and viral (imiquimod (3g/mL), R848 (1.5g/mL), and poly I:C (10g/mL)) surrogates or live rhinovirus (RV) 16 (MOI1). Cell-free supernatant was collected after 1h for neutrophil elastase (NE) and matrix metalloproteinase- (MMP-) 9 measurements or after 24h for CXCL8 release. Results. Constitutive NE was enhanced in AGood neutrophils compared to HC. fMLF stimulated neutrophils from ASubopt but not AGood produced 50% of HC levels. fMLF induced MMP-9 was impaired in ASubopt and AGood compared to HC. fMLF stimulated CXCL8 but not MMP-9 was positively correlated with FEV1 and FEV1/FVC. ASubopt and AGood responded similarly to other stimuli.Circulating neutrophils are different in asthma; however, this is likely to be related to airflow limitation rather than asthma control.
Grafton, K., Moir, L., Black, J., Hansbro, N., Hansbro, P., Burgess, J. & Oliver, B.G. 2014, 'LF-15 & T7, Synthetic Peptides Derived from Tumstatin, Attenuate Aspects of Airway Remodelling in a Murine Model of Chronic OVA-Induced Allergic Airway Disease', Plos One, vol. 9, no. 1.
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Background: Tumstatin is a segment of the collagen-IV protein that is markedly reduced in the airways of asthmatics. Tumstatin can play an important role in the development of airway remodelling associated with asthma due to its anti-angiogenic propertie
Jaffar, J., Unger, S., Corte, T.J., Keller, M., Wolters, P.J., Richeldi, L., Cerri, S., Prêle, C.M., Hansbro, P.M., Argraves, W.S., Oliver, R.A., Oliver, B.G., Black, J.L. & Burgess, J.K. 2014, 'Fibulin-1 predicts disease progression in patients with idiopathic pulmonary fibrosis.', Chest, vol. in press.
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Background The underlying mechanisms of idiopathic pulmonary fibrosis (IPF) are unknown. This progressive disease has high mortality rates and current models for prediction of mortality have limited value in identifying which patients will progress. We previously showed that the glycoprotein fibulin-1 is involved in enhanced proliferation and wound repair by mesenchymal cells, thus may contribute to lung fibrosis in IPF. Methods Serum, lung tissue and lung function values were obtained from four independent locations (Sydney and Perth, Australia, San Francisco, USA and Modena, Italy). Patients with IPF were followed for a minimum of one year and progression was defined as a significant decline in lung function or death. Primary parenchymal lung fibroblasts of 15 patients with and without IPF were cultured under non-stimulatory conditions. Fibulin-1 levels in serum and secreted or deposited by fibroblasts were measured by western blot and in lung tissue by immunohistochemistry. Results Serum fibulin-1 levels were increased in patients with IPF compared to subjects without lung disease (p=0.006). Furthermore, tissue fibulin-1 levels were increased in patients with IPF (p=0.02) and correlated negatively with lung function (r=-0.9, p<0.05). Primary parenchymal fibroblasts from patients with IPF produced more fibulin-1 than those from subjects without IPF (p<0.05). Finally, serum fibulin-1 levels at first blood draw predicted disease progression in IPF within 1 year (AUC 0.71, 95%CI 0.57 0.86, p=0.012). Conclusions Fibulin-1 is a novel potential biomarker for disease progression IPF and raise the possibility that it could be used as a target for the development of new treatments.
Chen, L., Ge, Q., Tjin, G., Alkhouri, H., Deng, L., Brandsma, C., Adcock, I., Timens, W., Postma, D.S., Burgess, J.K., Black, J.L. & Oliver, B.G. 2014, 'Effects of cigarette smoke extract on human airway smooth muscle cells in COPD', European Respiratory Journal, vol. 44, no. 3, pp. 634-646.
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We hypothesised that the response to cigarette smoke in airway smooth muscle (ASM) cells from smokers with chronic obstructive pulmonary disease (COPD) would be intrinsically different from smokers without COPD, producing greater pro-inflammatory mediators and factors relating to airway remodelling. ASM cells were obtained from smokers with or without COPD, and then stimulated with cigarette smoke extract (CSE) or transforming growth factor-1. The production of chemokines and matrix metalloproteinases (MMPs) were measured by ELISA, and the deposition of collagens by extracellular matrix ELISA. The effects of CSE on cell attachment and wound healing were measured by toluidine blue attachment and cell tracker green wound healing assays. CSE increased the release of CXCL8 and CXCL1 from human ASM cells, and cells from smokers with COPD produced more CSE-induced CXCL1. The production of MMP-1, -3 and -10, and the deposition of collagen VIII alpha 1 (COL8A1) were increased by CSE, especially in the COPD group which had higher production of MMP-1 and deposition of COL8A1. CSE decreased ASM cell attachment and wound healing in the COPD group only. ASM cells from smokers with COPD were more sensitive to CSE stimulation, which may explain, in part, why some smokers develop COPD.
Banville, N., Burgess, J.K., Jaffar, J., Tjin, G., Richeldi, L., Cerri, S., Persiani, E., Black, J.L. & Oliver, B.G. 2014, 'A Quantitative Proteomic Approach to Identify Significantly Altered Protein Networks in the Serum of Patients with Lymphangioleiomyomatosis (LAM)', PLoS One, vol. 9, no. 8.
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Lymphangioleiomyomatosis (LAM) is a rare and progressive cystic lung condition affecting approximately 3.47.5/million women, with an average lag time between symptom onset and diagnosis of upwards of 4 years. The aim of this work was to identify altered proteins in LAM serum which may be potential biomarkers of disease. Serum from LAM patient volunteers and healthy control volunteers were pooled and analysis carried out using quantitative 4-plex iTRAQ technology. Differentially expressed proteins were validated using ELISAs and pathway analysis was carried out using Ingenuity Pathway Analysis. Fourteen proteins were differentially expressed in LAM serum compared to control serum (p<0.05). Further screening validated the observed differences in extracellular matrix remodelling proteins including fibronectin (30% decrease in LAM, p = 0.03), von Willebrand Factor (40% reduction in LAM, p = 0.03) and Kallikrein III (25% increase in LAM, p = 0.03). Pathway networks elucidated the relationships between the ECM and cell trafficking in LAM. This study was the first to highlight an imbalance in networks important for remodelling in LAM, providing a set of novel potential biomarkers. These understandings may lead to a new effective treatment for LAM in the future.
Alkhouri, H., Rumzhum, N., Rahman, M., FitzPatrick, M., De Pedro, M., Oliver, B.G., Bourke, J. & Ammit, A. 2014, 'TLR2 activation causes tachyphylaxis to 2-agonists in vitro and ex vivo: modelling bacterial exacerbation', Allergy, vol. 69, no. 9, pp. 1215-1222.
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Background Asthma is a widespread chronic health problem exacerbated by common viral and bacterial infections. Further research is required to understand how infection worsens asthma control in order to advance therapeutic options in the future. Recent research has revealed that 2-adrenergic receptor (2-AR) agonists lose bronchodilatory efficacy because the receptor-mediated molecular pathways responsible for their beneficial actions are desensitized by infection. To date, most studies have focussed on viral infection, leaving the impact of bacterial infection on 2-AR desensitization relatively under-investigated. We address this in this study. Methods and Results Utilizing an in vitro model of bacterial exacerbation in airway smooth muscle (ASM) cells, we show that activation of toll-like receptor 2 (TLR2; mimicking bacterial infection) in the presence of an inflammatory stimulus leads to 2-AR desensitization. This occurs via TLR2-dependent upregulation of cyclooxygenase 2 (COX-2) mRNA expression and increased secretion of PGE2. Importantly, PGE2 causes heterologous 2-AR desensitization and reduces cAMP production in response to short-acting (salbutamol) and long-acting (formoterol) 2-agonists. Thus, bacterial infectious stimuli act in a PGE2-dependent manner to severely curtail the beneficial actions of 2-agonists. The impact of 2-AR desensitization is demonstrated by reduced gene expression of the critical anti-inflammatory molecule MKP-1 in response to 2-agonists, as well as impaired bronchodilation in a mouse lung slices. Conclusions Taken together, our results show that, like viruses, bacteria induce prostanoid-dependent 2-AR desensitization on ASM cells. Notably, COX-2 inhibition with the specific inhibitor celecoxib represses PGE2 secretion, presenting a feasible pharmacological option for treatment of infectious exacerbation in asthma in the future.
Oliver, B.G., Robinson, P., Peters, M. & Black, J. 2014, 'Viral infections and asthma: an inflammatory interface?', The European respiratory journal, vol. 44, no. 6, pp. 1666-1681.
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Asthma is a chronic inflammatory disease of the airways in which the majority of patients respond to treatment with corticosteroids and -adrenoceptor agonists. Acute exacerbations of asthma substantially contribute to disease morbidity, mortality and healthcare costs, and are not restricted to patients who are not compliant with their treatment regimens. Given that respiratory viral infections are the principal cause of asthma exacerbations, this review article will explore the relationship between viral infections and asthma, and will put forward hypotheses as to why virus-induced exacerbations occur. Potential mechanisms that may explain why current therapeutics do not fully inhibit virus-induced exacerbations, for example, -adrenergic desensitisation and corticosteroid insensitivity, are explored, as well as which aspects of virus-induced inflammation are likely to be attenuated by current therapy.
Herbert, C., Zeng, Q.X., Shanmugasundaram, R., Garthwaite, L., Oliver, B.G. & Kumar, R.K. 2014, 'Response of airway epithelial cells to double-stranded RNA in an allergic environment.', Translational respiratory medicine, vol. 2, no. 1, p. 11.
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Respiratory viral infections are the most common trigger of acute exacerbations in patients with allergic asthma. The anti-viral response of airway epithelial cells (AEC) may be impaired in asthmatics, while cytokines produced by AEC may drive the inflammatory response. We investigated whether AEC cultured in the presence of Th2 cytokines associated with an allergic environment exhibited altered responses to double-stranded RNA, a virus-like stimulus.We undertook preliminary studies using the MLE-12 cell line derived from mouse distal respiratory epithelial cells, then confirmed and extended our findings using low-passage human AEC. Cells were cultured in the absence or presence of the Th2 cytokines IL-4 and IL-13 for 48&nbsp;hours, then stimulated with poly I:C for 4&nbsp;hours. Expression of relevant anti-viral response and cytokine genes was assessed by quantitative real-time PCR. Secretion of cytokine proteins was assessed by immunoassay.Following stimulation with poly I:C, MLE-12 cells pre-treated with Th2 cytokines exhibited significantly higher levels of expression of mRNA for the cytokine genes Cxcl10 and Cxcl11, as well as a trend towards increased expression of Cxcl9 and Il6. Expression of anti-viral response genes was mostly unchanged, although Stat1, Ifit1 and Ifitm3 were significantly increased in Th2 cytokine pre-treated cells. Human AEC pre-treated with IL-4 and IL-13, then stimulated with poly I:C, similarly exhibited significantly higher expression of IL8, CXCL9, CXCL10, CXCL11 and CCL5 genes. In parallel, there was significantly increased secretion of CXCL8 and CCL5, as well as a trend towards increased secretion of CXCL10 and IL-6. Again, expression of anti-viral response genes was not decreased. Rather, there was significantly enhanced expression of mRNA for type III interferons, RNA helicases and other interferon-stimulated genes.The Th2 cytokine environment appears to promote increased production of pro-inflammatory chemokines by AEC in response to do...
Faiz, A., Tjin, G., Harkness, L., Weckmann, M., Bao, S., Black, J.L., Oliver, B.G.G. & Burgess, J.K. 2014, 'Correction: The expression and activity of cathepsins D, H and K in asthmatic airways', PLoS ONE, vol. 9, no. 1.
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Krimmer, D., Ichimaru, Y., Burgess, J., Black, J. & Oliver, B.G. 2013, 'Exposure to Biomass Smoke Extract Enhances Fibronectin Release from Fibroblasts', Plos One, vol. 8, no. 12.
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COPD induced following biomass smoke exposure has been reported to be associated with a more fibrotic phenotype than cigarette smoke induced COPD. This study aimed to investigate if biomass smoke induced extracellular matrix (ECM) protein production from
Keglowich, L., Roth, M., Philippova, M., Resink, T., Tjin, G., Oliver, B.G., Lardinois, D., Dessus-babus, S., Gosens, R., Haack, K., Tamm, M. & Borger, P. 2013, 'Bronchial Smooth Muscle Cells of Asthmatics Promote Angiogenesis through Elevated Secretion of CXC-Chemokines (ENA-78, GRO-alpha, and IL-8)', Plos One, vol. 8, no. 12.
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Background: Airway wall remodelling is a key pathology of asthma. It includes thickening of the airway wall, hypertrophy and hyperplasia of bronchial smooth muscle cells (BSMC), as well as an increased vascularity of the sub-epithelial cell layer. BSMC a
Van Ly, D., De Pedro, M., James, P., Morgan, L., Black, J., Burgess, J. & Oliver, B.G. 2013, 'Inhibition of phosphodiesterase 4 modulates cytokine induction from toll like receptor activated, but not rhinovirus infected, primary human airway smooth muscle', Respiratory Research, vol. 14.
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Background: Virus-induced exacerbations of Chronic Obstructive Pulmonary Disease (COPD) are a significant health burden and occur even in those receiving the best current therapies. Rhinovirus (RV) infections are responsible for half of all COPD exacerba
Hirota, J., Im, H., Rahman, M., Rumzhum, N., Manetsch, M., Pascoe, C., Bunge, K., Alkhouri, H., Oliver, B.G. & Ammit, A. 2013, 'The Nucleotide-Binding Domain and Leucine-Rich Repeat Protein-3 Inflammasome Is Not Activated in Airway Smooth Muscle Upon Toll-Like Receptor-2 Ligation', American Journal Of Respiratory Cell And Molecular Biology, vol. 49, no. 4, pp. 517-524.
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Inflammasomes have emerged as playing key roles in inflammation and innate immunity. A growing body of evidence has suggested that the nucleotide-binding domain and leucine-rich repeat protein-3 (NLRP3) inflammasome is important in chronic airway disease
Chen, L., Ge, Q., Black, J., Deng, L., Burgess, J. & Oliver, B.G. 2013, 'Differential Regulation of Extracellular Matrix and Soluble Fibulin-1 Levels by TGF-beta(1) in Airway Smooth Muscle Cells', Plos One, vol. 8, no. 6.
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Fibulin-1 (FBLN-1) is a secreted glycoprotein that is associated with extracellular matrix (ECM) formation and rebuilding. Abnormal and exaggerated deposition of ECM proteins is a hallmark of many fibrotic diseases, such as chronic obstructive pulmonary
Faiz, A., Tjin, G., Harkness, L., Weckmann, M., Bao, S., Black, J., Oliver, B.G. & Burgess, J. 2013, 'The Expression and Activity of Cathepsins D, H and K in Asthmatic Airways', Plos One, vol. 8, no. 3.
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Tumstatin is an anti-angiogenic collagen IV alpha 3 fragment, levels of which are reduced in the airways of asthmatics. Its reduction may be due to the degradation by extracellular matrix (ECM) proteases. Cathepsins play a role in ECM remodelling, with c
Van Ly, D., Faiz, A., Jenkins, C., Crossett, B., Black, J., Mcparland, B., Burgess, J. & Oliver, B.G. 2013, 'Characterising the Mechanism of Airway Smooth Muscle beta(2) Adrenoceptor Desensitization by Rhinovirus Infected Bronchial Epithelial Cells', Plos One, vol. 8, no. 2.
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Rhinovirus (RV) infections account for approximately two thirds of all virus-induced asthma exacerbations and often result in an impaired response to beta 2 agonist therapy. Using an in vitro model of RV infection, we investigated the mechanisms underlyi
Yeganeh, B., Xia, C., Movassagh, H., Koziol-white, C., Chang, Y., Al-alwan, L., Bourke, J. & Oliver, B.G. 2013, 'Emerging mediators of airway smooth muscle dysfunction in asthma', Pulmonary Pharmacology & Therapeutics, vol. 26, no. 1, pp. 105-111.
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Phenotypic changes in airway smooth muscle are integral to the pathophysiological changes that constitute asthma namely inflammation, airway wall remodelling and bronchial hyperresponsiveness. In vitro and in vivo studies have shown that the proliferativ
Im, H., Hirota, J., Rahman, M., Rumzhum, N., Manetsch, M., Pascoe, C., Oliver, B.G. & Ammit, A. 2013, 'The NLRP3 inflammasome is not activated in airway smooth muscle upon TLR2 ligation', European Respiratory Journal, vol. 42, p. 559.
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Inflammasomes have emerged as playing key roles in inflammation and innate immunity. A growing body of evidence has suggested that the nucleotide-binding domain and leucine-rich repeat protein 3 (NLRP3) inflammasome is important in chronic airway diseases such as asthma and COPD. Inflammasome activation results, in part, in pro-IL-1 processing and secretion of the pro-inflammatory cytokine IL-1. Because asthma exacerbations are associated with elevated levels of secreted IL-1 we addressed whether the NLRP3 inflammasome is activated under in vitro conditions that mimic infectious exacerbation in asthma. Primary cultures of airway smooth muscle (ASM) cells were treated with infectious stimuli (mimicked using the TLR2 agonist Pam3CSK4, a synthetic bacterial lipopeptide). While Pam3CSK4 robustly upregulated ASM cytokine expression in response to TNFa and significantly enhanced IL-1 mRNA expression, we were unable to detect IL-1 in the cell supernatants. Thus, IL-1 was not secreted and therefore unable to act in an autocrine manner to promote amplification of ASM inflammatory responses. Moreover, TLR2 ligation did not enhance NLRP3 mRNA expression in ASM cells, nor was NLRP3 protein detected in the airway smooth muscle layer of tracheal sections from human donors. In conclusion, these data demonstrate that enhanced synthetic function of ASM cells, induced by infectious exacerbation of airway inflammation, is NLRP3 inflammasome and IL-1-independent. Activation of NLRP3 inflammasome by invading pathogens may prove cell-type specific in exacerbation of airway inflammation in asthma.
Tan, X., Alrashdan, Y.A., Alkhouri, H., Oliver, B.G., Armour, C.L. & Hughes, J.M. 2013, 'Airway smooth muscle CXCR3 ligand production: regulation by JAK-STAT1 and intracellular Ca2+', American Journal of Physiology - Lung Cellular and Molecular Physiology, vol. 304, no. 11, pp. 790-802.
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In asthma, airway smooth muscle (ASM) chemokine (C-X-C motif) receptor 3 (CXCR3) ligand production may attract mast cells or T lymphocytes to the ASM, where they can modulate ASM functions. In ASM cells (ASMCs) from people with or without asthma, we aimed to investigate JAK-STAT1, JNK, and Ca2+ involvement in chemokine (C-X-C motif) ligand (CXCL)10 and CXCL11 production stimulated by interferon-?, IL-1, and TNF-a combined (cytomix). Confluent, growth-arrested ASMC were treated with inhibitors for pan-JAK (pyridone-6), JAK2 (AG-490), JNK (SP-600125), or the sarco(endo)plasmic reticulum Ca2+ATPase (SERCA) pump (thapsigargin), Ca2+ chelator (BAPTA-AM), or vehicle before and during cytomix stimulation for up to 24 h. Signaling protein activation as well as CXCL10/CXCL11 mRNA and protein production were examined using immunoblot analysis, real-time PCR, and ELISA, respectively. Cytomix-induced STAT1 activation was lower and CXCR3 ligand mRNA production was more sensitive to pyridone-6 and AG-490 in asthmatic than nonasthmatic ASMCs, but CXCL10/CXCL11 release was inhibited by the same proportion. Neither agent caused additional inhibition of release when used in combination with the JNK inhibitor SP-600125. Conversely, p65 NF-?B activation was higher in asthmatic than nonasthmatic ASMCs. BAPTA-AM abolished early CXCL10/CXCL11 mRNA production, whereas thapsigargin reduced it in asthmatic cells and inhibited CXCL10/CXCL11 release by both ASMC types. Despite these inhibitory effects, neither Ca2+ agent affected early activation of STAT1, JNK, or p65 NF-?B. In conclusion, intracellular Ca2+ regulated CXCL10/CXCL11 production but not early activation of the signaling molecules involved. In asthma, reduced ASM STAT1-JNK activation, increased NF-?B activation, and altered Ca2+ handling may contribute to rapid CXCR3 ligand production and enhanced inflammatory cell recruitment.
Beckett, E.L., Stevens, R.L., Jarnicki, A.G., Kim, R.Y., Hanish, I., Hansbro, N.G., Deane, A., Keely, S., Horvat, J.C., Yang, M., Oliver, B.G., van Rooijen, N., Inman, M.D., Adachi, R., Soberman, R.J., Hamadi, S., Wark, P.A., Foster, P.S. & Hansbro, P.M. 2013, 'A new short-term mouse model of chronic obstructive pulmonary disease identifies a role for mast cell tryptase in pathogenesis.', The Journal of allergy and clinical immunology, vol. 131, no. 3, pp. 752-762.
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Cigarette smoke-induced chronic obstructive pulmonary disease (COPD) is a life-threatening inflammatory disorder of the lung. The development of effective therapies for COPD has been hampered by the lack of an animal model that mimics the human disease in a short timeframe.We sought to create an early-onset mouse model of cigarette smoke-induced COPD that develops the hallmark features of the human condition in a short time-frame. We also sought to use this model to better understand pathogenesis and the roles of macrophages and mast cells (MCs) in patients with COPD.Tightly controlled amounts of cigarette smoke were delivered to the airways of mice, and the development of the pathologic features of COPD was assessed. The roles of macrophages and MC tryptase in pathogenesis were evaluated by using depletion and in vitro studies and MC protease 6-deficient mice.After just 8 weeks of smoke exposure, wild-type mice had chronic inflammation, mucus hypersecretion, airway remodeling, emphysema, and reduced lung function. These characteristic features of COPD were glucocorticoid resistant and did not spontaneously resolve. Systemic effects on skeletal muscle and the heart and increased susceptibility to respiratory tract infections also were observed. Macrophages and tryptase-expressing MCs were required for the development of COPD. Recombinant MC tryptase induced proinflammatory responses from cultured macrophages.A short-term mouse model of cigarette smoke-induced COPD was developed in which the characteristic features of the disease were induced more rapidly than in existing models. The model can be used to better understand COPD pathogenesis, and we show a requirement for macrophages and tryptase-expressing MCs.
Weckmann, M., Moir, L., Heckman, C., Black, J., Oliver, B.G. & Burgess, J. 2012, 'Lamstatin - a novel inhibitor of lymphangiogenesis derived from collagen IV', Journal of Cellular and Molecular Medicine, vol. 16, no. 12, pp. 3062-3073.
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The lymphatic system is essential for the maintenance of tissue homeostasis and immunity. Its dysfunction in disease (such as lymphangioleiomyomatosis) can lead to chylous effusions, oedema or dissemination of malignant cells. Collagen IV has six a chain
Ge, Q., Moir, L., Trian, T., Niimi, K., Poniris, M., Shepherd, P., Black, J., Oliver, B.G. & Burgess, J. 2012, 'The phosphoinositide 3 '-kinase p110 delta modulates contractile protein production and IL-6 release in human airway smooth muscle', Journal of Cellular Physiology, vol. 227, no. 8, pp. 3044-3052.
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Transforming growth factor (TGF) beta 1 increases pro-inflammatory cytokines and contractile protein expression by human airway smooth muscle (ASM) cells, which could augment airway inflammation and hyperresponsiveness. Phosphoinositide 3' kinase (PI3K)
Essilfie, A., Simpson, J., Dunkley, M., Morgan, L., Oliver, B.G., Gibson, P., Foster, P. & Hansbro, P. 2012, 'Combined Haemophilus influenzae respiratory infection and allergic airways disease drives chronic infection and features of neutrophilic asthma', Thorax, vol. 67, no. 7, pp. 588-599.
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Background 20-30% of patients with asthma have neutrophilic airway inflammation and reduced responsiveness to steroid therapy. They often have chronic airway bacterial colonisation and Haemophilus influenzae is one of the most commonly isolated bacteria.
Tan, X., Khalil, N., Tesarik, C., Vanapalli, K., Yaputra, V., Alkhouri, H., Oliver, B.G., Armour, C. & Hughes, J. 2012, 'Th1 cytokine-induced syndecan-4 shedding by airway smooth muscle cells is dependent on mitogen-activated protein kinases', American Journal Of Physiology-Lung Cellular And Molecular Physiology, vol. 302, no. 7, pp. 1700-1710.
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Tan X, Khalil N, Tesarik C, Vanapalli K, Yaputra V, Alkhouri H, Oliver BG, Armour CL, Hughes JM. Th1 cytokine-induced syndecan-4 shedding by airway smooth muscle cells is dependent on mitogen-activated protein kinases. Am J Physiol Lung Cell Mol Physiol
Baraket, M., Oliver, B.G., Burgess, J., Lim, S., King, G. & Black, J. 2012, 'Is low dose inhaled corticosteroid therapy as effective for inflammation and remodeling in asthma? A randomized, parallel group study', Respiratory Research, vol. 13.
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Background: While most of the clinical benefits of inhaled corticosteroid (ICS) therapy may occur at low doses, results of dose-ranging studies are inconsistent. Although symptom/lung function response to low and high dose ICS medication is comparable, i
Krimmer, D., Burgess, J.K., Wooi, T.K., Black, J.L. & Oliver, B.G. 2012, 'Matrix Proteins from Smoke-Exposed Fibroblasts Are Pro-proliferative', American Journal of Respiratory Cell and Molecular Biology, vol. 46, no. 1, pp. 34-39.
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Airway remodeling decreases lung function in chronic obstructive pulmonary disease (COPD). Extracellular matrix (ECM) deposition is increased in remodeled airways and drives cellular processes of proliferation, migration, and inflammation. We investigated the role of cigarette smoke in altering the ECM deposited from human lung fibroblasts. Lung fibroblasts isolated from patients with COPD or other lung disease were exposed to cigarette smoke extract (CSE) and 5 ng/ml transforming growth factor-1 for 72 hours; in some experiments, inhibitors of signaling molecules were added. Deposition of perlecan, fibronectin, and elastin were measured by ELISA, as was release of IL-8 and IL-13. Unstimulated fibroblast cells were reseeded onto deposited matrix and assessed for proliferation and cytokine release. CSE (5%) increased deposition of fibronectin and perlecan from only COPD fibroblasts. Fibronectin and perlecan deposition was attenuated by addition of the NF-?B inhibitor, BMS-345541, and the signal transduction and activator of transcription-1/3 inhibitor, pyridone 6, respectively. CSE (5%) increased IL-8 release from COPD fibroblasts more than non-COPD fibroblasts. This increase was attenuated by BMS-345541. Matrix deposited after 5% CSE stimulation increased proliferation of fibroblasts, but did not alter cytokine release. ECM produced from COPD fibroblasts after CSE exposure has proproliferative effects. Thus, the ECM in patients with COPD may create an environment that promotes airway remodeling.
Ichimaru, Y., Krimmer, D., Burgess, J.K., Black, J.L. & Oliver, B.G. 2012, 'TGF-beta enhances deposition of perlecan from COPD airway smooth muscle', American Journal of Physiology - Lung Cellular and Molecular Physiology, vol. 302, no. 3, pp. 325-333.
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Chronic obstructive pulmonary disease (COPD) and asthma are characterized by irreversible remodeling of the airway walls, including thickening of the airway smooth muscle layer. Perlecan is a large, multidomain, proteoglycan that is expressed in the lungs, and in other organ systems, and has been described to have a role in cell adhesion, angiogenesis, and proliferation. This study aimed to investigate functional properties of the different perlecan domains in relation to airway smooth muscle cells (ASMC). Primary human ASMC obtained from donors with asthma (n = 13), COPD (n = 12), or other lung disease (n = 20) were stimulated in vitro with 1 ng/ml transforming growth factor-1 (TGF-1) before perlecan deposition and cytokine release were analyzed. In some experiments, inhibitors of signaling molecules were added. Perlecan domains IV were seeded on tissue culture plates at 10 &micro;g/ml with 1 &micro;g/ml collagen I as a control. ASM was incubated on top of the peptides before being analyzed for attachment, proliferation, and wound healing. TGF-1 upregulated deposition of perlecan by ASMC from COPD subjects only. TGF-1 upregulated release of IL-6 into the supernatant of ASMC from all subjects. Inhibitors of SMAD and JNK signaling molecules decreased TGF-1-induced perlecan deposition by COPD ASMC. Attachment of COPD ASMC was upregulated by collagen I and perlecan domains IV and V, while perlecan domain II upregulated attachment only of asthmatic ASMC. Seeding on perlecan domains did not increase proliferation of any ASMC type. TGF-1-induced perlecan deposition may enhance attachment of migrating ASMC in vivo and thus may be a mechanism for ASMC layer hypertrophy in COPD.
Ge, Q., Moir, L.M., Trian, T., Niimi, K., Poniris, M., Shepherd, P.R., Black, J.L., Oliver, B.G. & Burgess, J.K. 2012, 'The phosphoinositide 3'-kinase p110d modulates contractile protein production and IL-6 release in human airway smooth muscle', Journal of Cellular Physiology, vol. 227, no. 8, pp. 3044-3052.
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Transforming growth factor (TGF) 1 increases pro-inflammatory cytokines and contractile protein expression by human airway smooth muscle (ASM) cells, which could augment airway inflammation and hyperresponsiveness. Phosphoinositide 3' kinase (PI3K) is one of the signaling pathways implicated in TGF1 stimulation, and may be altered in asthmatic airways. This study compared the expression of PI3K isoforms by ASM cells from donors with asthma (A), chronic obstructive pulmonary disease (COPD), or neither disease (NA), and investigated the role of PI3K isoforms in the production of TGF1 induced pro-inflammatory cytokine and contractile proteins in ASM cells. A cells expressed higher basal levels of p110d mRNA compared to NA and COPD cells; however COPD cells produced more p110d protein. TGF1 increased 110d mRNA expression to the same extent in the three groups. Neither the p110d inhibitor IC87114 (1, 10, 30?&micro;M), the p110 inhibitor TGX221 (0.1, 1, 10?&micro;M) nor the PI3K pan inhibitor LY294002 (3, 10?&micro;M) had any effect on basal IL-6, calponin or smooth muscle a-actin (a-SMA) expression. However, TGF1 increased calponin and a-SMA expression was inhibited by IC87114 and LY294002 in all three groups. IC87114, TGX221, and LY294002 reduced TGF1 induced IL-6 release in a dose related manner in all groups of ASM cells. PI3K p110d is important for TGF1 induced production of the contractile proteins calponin and a-SMA and the proinflammatory cytokine IL-6 in ASM cells, and may therefore be relevant as a potential therapeutic target to treat both inflammation and airway remodeling.
Niimi, K., Ge, Q., Moir, L.M., Ammit, A., Trian, T., Burgess, J.K., Black, J.L. & Oliver, B.G. 2012, 'Beta2-Agonists upregulate PDE4 mRNA but not protein or activity in human airway smooth muscle cells from asthmatic and nonasthmatic volunteers', American Journal of Physiology - Lung Cellular and Molecular Physiology, vol. 302, no. 3, pp. 334-342.
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2-Adrenergic receptor (2AR) agonists induce airway relaxation via cAMP. Phosphodiesterase (PDE)s degrade and regulate cAMP, and in airway smooth muscle (ASM) cells PDE4D degrades cAMP. Long-acting 2-agonists are now contraindicated as monotherapy for asthma, and increased PDE4D has been speculated to contribute to this phenomenon. In this study we investigated the expression of PDE4D in asthmatic and nonasthmatic ASM cells and its regulation by formoterol and budesonide. Primary ASM cells from people with or without asthma were stimulated with transforming growth factor (TGF)-1, formoterol, and/or budesonide. PDE4D mRNA was assessed by real-time PCR, or PCR to assess splice variant production. PDE4D protein was assessed by Western blotting, and we investigated the effect of formoterol on cAMP production and PDE activity. Interleukin (IL)-6 was assessed using ELISA. PDE4D mRNA was dose dependently upregulated by formoterol, with a single splice variant, PDE4D5, present. Formoterol did not induce PDE4D protein at time points between 3 to 72 h, whereas it did induce and increase IL-6 secretion. We pretreated cells with actinomycin D and a proteasome inhibitor, MG132, and found no evidence of alterations in mRNA, protein expression, or degradation of PDE4D. Finally PDE activity was not altered by formoterol. This study shows, for the first time, that PDE4D5 is predominantly expressed in human ASM cells from people with and without asthma and that formoterol does not upregulate PDE4D protein production. This leads us to speculate that continual therapy with 2AR agonists is unlikely to cause PDE4-mediated tachyphylaxis.
Black, J.L., Burgess, J. & Oliver, B. 2012, 'On the one hand, on the other hand', Journal of Applied Physiology, vol. 113, no. 5, p. 845.
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Kuo, C., Lim, S., King, N.J.C., Bartlett, N.W., Walton, R.P., Zhu, J., Glanville, N., Aniscenko, J., Johnston, S.L., Burgess, J.K., Black, J.L. & Oliver, B.G. 2012, 'Erratum: Rhinovirus infection induces expression of airway remodelling factors in vitro and in vivo (Respirology (2011) 16 (367-377))', Respirology, vol. 17, no. 1, p. 192.
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Van Ly, D., Burgess, J.K., Brock, T.G., Lee, T.H., Black, J.L. & Oliver, B.G.G. 2012, 'Prostaglandins but not leukotrienes alter extracellular matrix protein deposition and cytokine release in primary human airway smooth muscle cells and fibroblasts.', American journal of physiology. Lung cellular and molecular physiology, vol. 303, no. 3, pp. L239-L250.
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Eicosanoids are lipid-signaling mediators released by many cells in response to various stimuli. Increasing evidence suggests that eicosanoids such as leukotrienes and prostaglandins (PGs) may directly mediate remodeling. In this study, we assessed whether these substances could alter extracellular matrix (ECM) proteins and the inflammatory profiles of primary human airway smooth muscle cells (ASM) and fibroblasts. PGE(2) decreased both fibronectin and tenascin C in fibroblasts but only fibronectin in ASM. PGD(2) decreased both fibronectin and tenascin C in both ASM and fibroblasts, whereas PGF(2) had no effect on ECM deposition. The selective PGI(2) analog, MRE-269, decreased fibronectin but not tenascin C in both cell types. All the PGs increased IL-6 and IL-8 release in a dose-dependent manner in ASM and fibroblasts. Changes in ECM deposition and cytokine release induced by prostaglandins in both ASM and fibroblasts were independent of an effect on cell number. Neither the acute nor repeated stimulation with leukotrienes had an effect on the deposition of ECM proteins or cytokine release from ASM or fibroblasts. We concluded that, collectively, these results provide evidence that PGs may contribute to ECM remodeling to a greater extent than leukotrienes in airway cells.
Bosse, Y., Vagula, M.C., Rawding, R.S., Pun, M., Black, J.L., Burgess, J., Oliver, B., Berger, P., Marthan, R. & Adner, M. 2012, 'Comments on Point:Counterpoint: Alterations in airway smooth muscle phenotype do/do not cause airway hyperresponsiveness in asthma.', Journal of applied physiology (Bethesda, Md. : 1985), vol. 113, no. 5, pp. 844-846.
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Borger, P., Oliver, B., Heijink, I. & Hardavella, G. 2012, 'Beyond the Immune System: The Role of Resident Cells in Asthma and COPD', Journal of Allergy, vol. 2012, pp. 1-3.
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Trian, T., Burgess, J., Niimi, K., Moir, L., Ge, Q., Berger, P., Liggett, S., Black, J. & Oliver, B.G. 2011, 'beta(2)-Agonist Induced cAMP Is Decreased in Asthmatic Airway Smooth Muscle Due to Increased PDE4D', Plos One, vol. 6, no. 5.
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Background and Objective: Asthma is associated with airway narrowing in response to bronchoconstricting stimuli and increased airway smooth muscle (ASM) mass. In addition, some studies have suggested impaired beta-agonist induced ASM relaxation in asthma
Moir, L., Trian, T., Ge, Q., Shepherd, P., Burgess, J., Oliver, B.G. & Black, J. 2011, 'Phosphatidylinositol 3-Kinase Isoform-Specific Effects in Airway Mesenchymal Cell Function', Journal Of Pharmacology And Experimental Therapeutics, vol. 337, no. 2, pp. 557-566.
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The phosphatidylinositol 3-kinase (PI3K) signal transduction pathway is implicated in the airway remodeling associated with asthma. The class IA PI3K isoforms are known to be activated by growth factors and cytokines. Because this pathway is a possible s
Kuo, C., Lim, S., King, N., Bartlett, N., Walton, R., Zhu, J., Glanville, N., Aniscenko, J., Johnston, S., Burgess, J., Black, J. & Oliver, B.G. 2011, 'Rhinovirus infection induces expression of airway remodelling factors in vitro and in vivo', Respirology, vol. 16, no. 2, pp. 367-377.
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Background and objective: A hallmark of asthma is airway remodelling, which includes increased deposition of extracellular matrix (ECM) protein. Viral infections may promote the development of asthma and are the most common causes of asthma exacerbations
Trian, T., Burgess, J.K., Niimi, K., Moir, L.M., Ge, Q., Berger, P., Liggett, S.B., Black, J.L. & Oliver, B.G. 2011, 'b2-Agonist Induced cAMP Is Decreased in Asthmatic Airway Smooth Muscle Due to Increased PDE4D', PLoS One, vol. 6, no. 5.
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Background and Objective: Asthma is associated with airway narrowing in response to bronchoconstricting stimuli and increased airway smooth muscle (ASM) mass. In addition, some studies have suggested impaired b-agonist induced ASM relaxation in asthmatics, but the mechanism is not known. Objective: To characterize the potential defect in b-agonist induced cAMP in ASM derived from asthmatic in comparison to non-asthmatic subjects and to investigate its mechanism. Methods: We examined b2-adrenergic (b2AR) receptor expression and basal b-agonist and forskolin (direct activator of adenylyl cyclase) stimulated cAMP production in asthmatic cultured ASM (n = 15) and non-asthmatic ASM (n = 22). Based on these results, PDE activity, PDE4D expression and cell proliferation were determined. Results: In the presence of IBMX, a pan PDE inhibitor, asthmatic ASM had ,50% lower cAMP production in response to isoproterenol, albuterol, formoterol, and forskolin compared to non-asthmatic ASM. However when PDE4 was specifically inhibited, cAMP production by the agonists and forskolin was normalized in asthmatic ASM. We then measured the amount and activity of PDE4, and found ,2-fold greater expression and activity in asthmatic ASM compared to non-asthmatic ASM. Furthermore, inhibition of PDE4 reduced asthmatic ASM proliferation but not that of non-asthmatic ASM. Conclusion: Decreased b-agonist induced cAMP in ASM from asthmatics results from enhanced degradation due to increased PDE4D expression. Clinical manifestations of this dysregulation would be suboptimal b-agonist-mediated bronchodilation and possibly reduced control over increasing ASM mass. These phenotypes appear to be ``hard-wired into ASM from asthmatics, as they do not require an inflammatory environment in culture to be observed.
Van Ly, D., King, N.J., Moir, L.M., Burgess, J.K., Black, J.L. & Oliver, B.G. 2011, 'Effects of 2 Agonists, Corticosteroids, and Novel Therapies on Rhinovirus-Induced Cytokine Release and Rhinovirus Replication in Primary Airway Fibroblasts', Journal of Allergy, vol. 2011.
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Rhinovirus-(RV-) induced asthma exacerbations account for high asthma-related health costs and morbidity in Australia. The cellular mechanism underlying this pathology is likely the result of RV-induced nuclear-factor-kappa-B-(NF-?B-) dependent inflammation. NF-?B may also be important in RV replication as inhibition of NF-?B inhibits replication of other viruses such as human immunodeficiency virus and cytomegalovirus. To establish the role of NF-?B inhibitors in RV-induced IL- 6 and IL-8 and RV replication, we used pharmacological inhibitors of NF-?B, and steroids and/or 2 agonists were used for comparison. Primary human lung fibroblasts were infected with RV-16 in the presence of NF-?B inhibitors: BAY-117085 and dimethyl fumarate; 2 agonist: salmeterol; and/or corticosteroids: dexamethasone; fluticasone. RV-induced IL-6 and IL-8 and RV replication were assessed using ELISAs and virus titration assays. RV replicated and increased IL-6 and IL-8 release. Salmeterol increased, while dexamethasone and fluticasone decreased RV-induced IL-6 and IL-8 (P < 0.05). The NF-?B inhibitor BAY-117085 inhibited only RV-induced IL-6 (P < 0.05) and dimethyl fumarate did not alter RV-induced IL-6 and IL-8. Dimethylfumarate increased RV replication whilst other drugs did not alter RV replication. These data suggest that inhibition of NF-?B alone is unlikely to be an effective treatment compared to current asthma therapeutics.
Krimmer, D. & Oliver, B.G. 2011, 'What can in vitro models of COPD tell us?', Pulmonary Pharmacology and Therapeutics, vol. 24, no. 5, pp. 471-477.
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Chronic obstructive pulmonary disease (COPD) is a progressive lung disease characterised by chronic bronchitis, largely irreversible remodelling of the small airways, and emphysematous destruction of the alveoli. COPD is projected to be the third leading cause of death worldwide by 2020. COPD often results from prolonged exposure to irritants such as cigarette smoke or inhaled particulates. Current pharmacotherapies for COPD are unable to reverse the pathological changes of this disease, and this is partially due to a limited understanding of the intricate mechanisms by which chronic exposure lead to the different pathological components of COPD. This review examines how the mechanisms that underlie various components of COPD can be modelled in vitro, specifically using cigarette smoke extract with cells cultured from primary human lung tissue, and how the effectiveness of current and novel pharmacotherapies on successfully attenuating these pathological changes can also be examined in vitro.
moir, L., ng, H., Poniris, M., santa, T., Burgess, J.K., Oliver, B.G., krymskaya, V. & Black, J.L. 2011, 'Doxycycline inhibits matrix metalloproteinase-2 secretion from TSC2-null mouse embryonic fibroblasts and lymphangioleiomyomatosis cells', British Journal Of Pharmacology, vol. 164, no. 1, pp. 83-92.
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BACKGROUND AND PURPOSE Lymphangioleiomyomatosis (LAM) is characterized by the abnormal growth of smooth muscle-like cells (LAM cells) and cystic destruction of the lung parenchyma. LAM cell-derived matrix metalloproteinases (MMPs) are thought to play a prominent role in the tissue destruction. The aim of this study was to determine whether doxycycline, a known MMP inhibitor, can inhibit LAM cell proliferation or mitochondrial function and/or modulate MMPs and their tissue inhibitors (TIMPs). EXPERIMENTAL APPROACH Wild-type and tuberous sclerosis complex-2 (TSC2)-null mouse embryonic fibroblasts (MEFs) were cultured in DMEM containing 10% fetal bovine serum (FBS). Human LAM cells were derived from the lungs of LAM patients and airway smooth muscle cells from control subjects. Cells were stimulated with FBS with or without doxycycline for up to 9 days. Proliferation was assessed by manual cell counts and MTT assay, MMP production by zymography and ELISA, and TIMP production using ELISA. KEY RESULTS Doxycycline did not change FBS-induced proliferation in MEFs or human cells. However, doxycycline did reduce metabolic activity of both wild-type and TSC2-null MEFs and LAM cells, but had no effect on control cells. Furthermore, doxycycline reduced MMP-2 from MEFs and decreased active-MMP-2 from LAM cells but had no effect on TIMP-1 and TIMP-2 from human LAM cells. CONCLUSIONS AND IMPLICATIONS Doxycycline decreased MMP levels and cell metabolic activity, which raises the possibility of therapeutic efficacy in LAM.
Kuo, C., Lim, S., King, N.J.C., Johnston, S.L., Burgess, J.K., Black, J.L. & Oliver, B.G. 2011, 'Rhinovirus infection induces extracellular matrix protein deposition in asthmatic and nonasthmatic airway smooth muscle cells.', American journal of physiology. Lung cellular and molecular physiology, vol. 300, no. 6, pp. L951-L957.
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Airway remodeling, which includes increases in the extracellular matrix (ECM), is a characteristic feature of asthma and is correlated to disease severity. Rhinovirus (RV) infections are associated with increased risk of asthma development in young children and are the most common cause of asthma exacerbations. We examined whether viral infections can increase ECM deposition and whether this increased ECM modulates cell proliferation and migration. RV infection of nonasthmatic airway smooth muscle (ASM) cells significantly increased the deposition of fibronectin (40% increase, n = 12) and perlecan (80% increase, n = 14), while infection of asthmatic ASM cells significantly increased fibronectin (75% increase, n = 9) and collagen IV (15% increase, n = 9). We then treated the ASM cells with the Toll-like receptor (TLR) agonists polyinosinic:polycytidylic acid, imiquimod, and pure RV RNA and were able to show that the mechanism through which RV induced ECM deposition was via the activation of TLR3 and TLR7/8. Finally, we assessed whether the virus-induced ECM was bioactive by measuring the amount of migration and proliferation of virus-naive cells that seeded onto the ECM. Basically, ECM from asthmatic ASM cells induced twofold greater migration of virus-naive ASM cells than ECM from nonasthmatic ASM cells, and these rates of migration were further increased on RV-modulated ECM. Increased migration on the RV-modulated ECM was not due to increased cell proliferation, as RV-modulated ECM decreased the proliferation of virus-naive cells. Our results suggest that viruses may contribute to airway remodeling through increased ECM deposition, which in turn may contribute to increased ASM mass via increased cell migration.
Tovey, E.R., Ng, D.S.Y., Stelzer-Braid, S., Rawlinson, W.D. & Oliver, B.G. 2011, 'Erratum: Children with asthma and no URTI, more commonly have rhinovirus in their exhaled breath, than in mucous (J Allergy Clin Immunol (2009) 123 (S171))', Journal of Allergy and Clinical Immunology, vol. 127, no. 2, p. 551.
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Ge, Q., Moir, L., Black, J., Oliver, B.G. & Burgess, J. 2010, 'TGF beta 1 Induces IL-6 and Inhibits IL-8 Release in Human Bronchial Epithelial Cells: The Role of Smad2/3', Journal of Cellular Physiology, vol. 225, no. 3, pp. 846-854.
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Human bronchial epithelial (HBE) cells contribute to asthmatic airway inflammation by secreting cytokines, chemokines, and growth factors, including interleukin (IL)-6, IL-8 and transforming growth factor (TGF) beta 1, all of which are elevated in asthma
Lau, J., Oliver, B.G., Baraket, M., Beckett, E., Hansbro, N., Moir, L., Wilton, S., Williams, C., Foster, P., Hansbro, P., Black, J. & Burgess, J. 2010, 'Fibulin-1 Is Increased in Asthma - A Novel Mediator of Airway Remodeling?', Plos One, vol. 5, no. 10.
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Background: The extracellular matrix is a dynamic and complex network of macromolecules responsible for maintaining and influencing cellular functions of the airway. The role of fibronectin, an extracellular matrix protein, is well documented in asthma.
Trian, T., Moir, L., Ge, Q., Burgess, J., Kuo, C., King, N., Reddel, H., Black, J., Oliver, B.G. & Mcparland, B. 2010, 'Rhinovirus-Induced Exacerbations of Asthma How Is the beta(2)-Adrenoceptor Implicated?', American Journal Of Respiratory Cell And Molecular Biology, vol. 43, no. 2, pp. 227-233.
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Rhinovirus (RV) infections are the major cause of asthma exacerbations in children and adults. Under normal circumstances, asthmatic airway obstruction improves spontaneously or characteristically briskly in response to inhaled beta(2)-adrenergic recepto
Lau, J., Oliver, B.G., Moir, L., Black, J. & Burgess, J. 2010, 'Differential expression of peroxisome proliferator activated receptor gamma and cyclin D1 does not affect proliferation of asthma- and non-asthma-derived airway smooth muscle cells', Respirology, vol. 15, no. 2, pp. 303-312.
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Background and objective: Airway remodelling involves thickening of the airway smooth muscle (ASM) bulk. Proliferation of asthma-derived ASM cells is increased in vitro, but underlying mechanisms remain unknown. Peroxisome proliferators activated recepto
Burgess, J., Boustany, S., Moir, L., Weckmann, M., Lau, J., Grafton, K., Baraket, M., Hansbro, P., Hansbro, N., Foster, P., Black, J. & Oliver, B.G. 2010, 'Reduction of Tumstatin in Asthmatic Airways Contributes to Angiogenesis, Inflammation, and Hyperresponsiveness', American Journal Of Respiratory And Critical Care Medicine, vol. 181, no. 2, pp. 106-115.
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Rationale Angiogenesis is a prominent feature of remodeling in asthma. Many proangiogenic factors are up-regulated in asthma, but little is known about levels of endogenous antiangiogenic agents. Collagen IV is decreased in the airway basement membrane i
Burgess, J.K., Boustany, S., Moir, L.M., Weckmann, M., Lau, J.Y., Grafton, K., Baraket, M., Hansbro, P.M., Hansbro, N.G., Foster, P.S., Black, J.L. & Oliver, B.G. 2010, 'Reduction of Tumstatin in Asthmatic Airways Contributes to Angiogenesis, Inflammation, and Hyperresponsiveness', American Journal of Respiratory and Critical Care Medicine, vol. 181, no. 2, pp. 106-115.
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Rationale: Angiogenesis is a prominent feature of remodeling in asthma. Many proangiogenic factors are up-regulated in asthma, but little is known about levels of endogenous antiangiogenic agents. Collagen IV is decreased in the airway basement membrane in asthma. It has six a chains, of which the noncollagenous domain-1 domains have endogenous antiangiogenic properties. Objectives: To study the expression of the noncollagenous domain-1 of the a3 chain of collagen IV, tumstatin, in the airways of subjects with and without asthma and to examine the potential for tumstatin to regulate angiogenesis and inflammation. Methods: We used immunohistochemistry and dot blots to examine the expression of tumstatin in bronchial biopsies, bronchoalveolar lavage fluid, and serum. We then used an in vitro angiogenesis assay and a murine model of allergic airways disease to explore tumstatin's biological function. Measurements and Main Results: The level of tumstatin is decreased 18-fold in the airways of patients with asthma but not in subjects without asthma, including those with chronic obstructive pulmonary disease, cystic fibrosis, and bronchiectasis. In vitro, recombinant tumstatin inhibited primary pulmonary endothelial cell tube formation. In a mouse model of chronic allergic airways disease, tumstatin suppressed angiogenesis, airway hyperresponsiveness, inflammatory cell infiltration, and mucus secretion and decreased levels of vascular endothelial growth factor and IL-13. Conclusions: The observation that tumstatin is decreased in asthmatic airways and inhibits airway hyperresponsiveness and angiogenesis demonstrates the potential use of antiangiogenic agents such as tumstatin as a therapeutic intervention in diseases that are characterized by aberrant angiogenesis and tissue remodeling, such as asthma.
Ge, Q., Moir, L.M., Black, J.L., Oliver, B.G. & Burgess, J.K. 2010, 'TGF1 induces IL-6 and inhibits IL-8 release in human bronchial epithelial cells: The role of Smad2/3', Journal of Cellular Physiology, vol. 225, no. 3, pp. 846-854.
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Human bronchial epithelial (HBE) cells contribute to asthmatic airway inflammation by secreting cytokines, chemokines, and growth factors, including interleukin (IL)-6, IL-8 and transforming growth factor (TGF) 1, all of which are elevated in asthmatic airways. This study examines the signaling pathways leading to TGF1 induced IL-6 and IL-8 in primary HBE cells from asthmatic and non-asthmatic volunteers. HBE cells were stimulated with TGF1 in the presence or absence of signaling inhibitors. IL-6 and IL-8 protein and mRNA were measured by ELISA and real-time PCR respectively, and cell signaling kinases by Western blot. TGF1 increased IL-6, but inhibited IL-8 production in both asthmatic and non-asthmatic cells; however, TGF induced significantly more IL-6 in asthmatic cells. Inhibition of JNK MAP kinase partially reduced TGF1 induced IL-6 in both cell groups. TGF1 induced Smad2 phosphorylation, and blockade of Smad2/3 prevented both the TGF1 modulated IL-6 increase and the decrease in IL-8 production in asthmatic and non-asthmatic cells. Inhibition of Smad2/3 also increased basal IL-8 release in asthmatic cells but not in non-asthmatic cells. Using CHIP assays we demonstrated that activated Smad2 bound to the IL-6, but not the IL-8 promoter region. We conclude that the Smad2/3 pathway is the predominant TGF1 signaling pathway in HBE cells, and this is altered in asthmatic bronchial epithelial cells. Understanding the mechanism of aberrant pro-inflammatory cytokine production in asthmatic airways will allow the development of alternative ways to control airway inflammation
Zhang, Y. & Oliver, B. 2010, 'An evolutionary consequence of dosage compensation on Drosophila melanogaster female X-chromatin structure?', BMC genomics, vol. 11, p. 6.
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BACKGROUND: X chromosomes are subject to dosage compensation in Drosophila males. Dosage compensation requires cis sequence features of the X chromosome that are present in both sexes by definition and trans acting factors that target chromatin modifying machinery to the X specifically in males. The evolution of this system could result in neutral X chromatin changes that will be apparent in females. RESULTS: We find that the general chromatin structure of female X chromosomes is distinct from autosomes. Additionally, specific histone marks associated with dosage compensation and active chromatin marks on the male X chromosome are also enriched on the X chromosomes of females, albeit to a lesser degree. CONCLUSIONS: Our data indicate that X chromatin structure is fundamentally different from autosome structure in both sexes. We suggest that the differences between the X chromosomes and autosomes in females are a consequence of mechanisms that have evolved to ensure sufficient X chromosome expression in the soma of males.
Black, J., Oliver, B.G. & Roth, M. 2009, 'Molecular Mechanisms of Combination Therapy With Inhaled Corticosteroids and Long-Acting beta-Agonists', Chest, vol. 136, no. 4, pp. 1095-1100.
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The treatment of asthma relies on the use of the following two major drug classes: beta(2)-agonists, both short acting and long acting; and corticosteroids (CSs). Although the properties of each drug class are well described, their use in combination del
Burgess, J., Ceresa, C., Johnson, S., Kanabar, V., Moir, L., Nguyen, T., Oliver, B.G., Schuliga, M. & Ward, J. 2009, 'Tissue and matrix influences on airway smooth muscle function', Pulmonary Pharmacology & Therapeutics, vol. 22, no. 5, pp. 379-387.
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Asthma is characterized by structural changes in the airways - airway remodelling. These changes include an increase in the bulk of the airway smooth muscle (ASM) and alterations in the profile of extracellular matrix (ECM) proteins in the airway wall. T
Stelzer-braid, S., Oliver, B.G., Blazey, A., Argent, E., Newsome, T., Rawlinson, W. & Tovey, E. 2009, 'Exhalation of Respiratory Viruses by Breathing, Coughing, and Talking', Journal of Medical Virology, vol. 81, no. 9, pp. 1674-1679.
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There is a lack of quantitative information about the generation of virus aerosols by infected subjects. The exhaled aerosols generated by coughing, talking, and breathing were sampled in 50 subjects using a novel mask, and analyzed using PCR for nine re
Fukuyama, S., Nakano, T., Matsumoto, T., Oliver, B.G., Burgess, J., Moriwaki, A., Tanaka, K., Kubo, M., Hoshino, T., Tanaka, H., Mckenzie, A., Matsumoto, K., Aizawa, H., Nakanishi, Y., Yoshimura, A., Black, J. & Inoue, H. 2009, 'Pulmonary Suppressor of Cytokine Signaling-1 Induced by IL-13 Regulates Allergic Asthma Phenotype', American Journal Of Respiratory And Critical Care Medicine, vol. 179, no. 11, pp. 992-998.
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Rationale: Th2 cytokines play an important role in allergic diseases. These cytokines activate signal transduction pathways, including Janus kinase/signal transducer and activator of transcription (STAT) signaling. Although the suppressor of cytokine sig
Krimmer, D., Loseli, M., Hughes, J.M., Oliver, B.G., Moir, L.M., Hunt, N.H., Black, J.L. & Burgess, J.K. 2009, 'CD40 and OX40 ligand are differentially regulated on asthmatic airway smooth muscle', Allergy, vol. 64, no. 7, pp. 1074-1082.
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Background: CD40 and OX40 Ligand (OX40L) are cell-surface molecules expressed on airway smooth muscle (ASM) that can enhance inflammatory cell activation and survival. The aim of this study was to examine the effect of tumour necrosis factor-alpha (TNF-a) and interferon-gamma (IFN-?) on ASM CD40 and OX40L expression. Methods: CD40 and OX40L expression on human ASM cells from asthmatic and nonasthmatic donors following stimulation with TNF-a and/or IFN-? was measured using cell-surface enzyme-linked immunosorbent assay (ELISA) and flow cytometry. Involvement of signalling pathway was investigated with pharmacological inhibitors. Soluble TNF receptor levels were quantified by ELISA. Results: Interferon-? and TNF-a synergistically increased CD40 expression to a greater extent on asthmatic than on nonasthmatic ASM. In contrast, IFN-? reduced TNF-a-induced OX40L expression to a similar extent in both cell types. TNF-a and IFN-? induced CD40 via nuclear factor-?B (NF-?B) and signal transducer and activator of transcription-3 in both cell types and modulated OX40L via NF-?B and c-Jun N terminal kinase in nonasthmatic cells. Similar effects on the induction of OX40L in asthmatic cells were seen with NF-?B, but these were not statistically significant. The reduced OX40L expression with TNF-a and IFN-? involved extracellular regulated kinase 1/2 activation. Conclusion: Asthmatic ASM may modulate airway inflammation locally by increasing CD40 and OX40L expression in response to cytokines. IFN-? may regulate ASM pro-inflammatory actions by differentially modulating ASM CD40 and OX40L expression.
p, S., merfort, I., Hughes, M.J., Oliver, B.G., Tamm, M. & Roth, M. 2009, 'Dimethylfumarate inhibits NFkB function at multiple levels to limit airway smooth muscle cell cytokine secretion', American Journal of Physiology - Lung Cellular and Molecular Physiology, vol. 297, no. 2, pp. 326-339.
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The antipsoriatic dimethylfumarate (DMF) has been anecdotically reported to reduce asthma symptoms and to improve quality of life of asthma patients. DMF decreases the expression of proinflammatory mediators by inhibiting the transcription factor NF-?B and might therefore be of interest for the therapy of inflammatory lung diseases. In this study, we determined the effect of DMF on platelet-derived growth factor (PDGF)-BB- and TNFa-induced asthma-relevant cytokines and NF-?B activation by primary human asthmatic and nonasthmatic airway smooth muscle cells (ASMC). Confluent nonasthmatic and asthmatic ASMC were incubated with DMF (0.1100 &micro;M) and/or dexamethasone (0.00010.1 &micro;M), NF-?B p65 siRNA (100 nM), the NF-?B inhibitor helenalin (1 &micro;M) before stimulation with PDGF-BB or TNFa (10 ng/ml). Cytokine release was measured by ELISA. NF-?B, mitogen and stress-activated kinase (MSK-1), and CREB activation was determined by immunoblotting and EMSA. TNFa-induced eotaxin, RANTES, and IL-6 as well as PDGF-BB-induced IL-6 expression was inhibited by DMF and by dexamethasone from asthmatic and nonasthmatic ASMC, but the combination of both drugs showed no glucocorticoid sparing effect in either of the two groups. NF-?B p65 siRNA and/or the NF-?B inhibitor helenalin reduced PDGF-BB- and TNFa-induced cytokine expression, suggesting the involvement of NF-?B signaling. DMF inhibited TNFa-induced NF-?B p65 phosphorylation, NF-?B nuclear entry, and NF-?B-DNA complex formation, whereas PDGF-BB appeared not to activate NF-?B within 60 min. Both stimuli induced the phosphorylation of MSK-1, NF-?B p65 at Ser276, and CREB, and all were inhibited by DMF. These data suggest that DMF downregulates cytokine secretion not only by inhibiting NF-?B but a wider range of NF-?B-linked signaling proteins, which may explain its potential beneficial effect in asthma.
Weckmann, M., Trian, T. & Oliver, B.G.G. 2009, 'Reconstruction is not renovation - The role of remodeling in asthma', Journal of Asthma and Allergy, no. 2, pp. 33-42.
The chronicity of asthma results not only in persistent lung inflammation but also in changes in structure and composition of this vital organ. These changes are most commonly referred to as remodeling, and include epithelial dysplasia, angiogenesis, changes in the extracellular matrix and increased smooth muscle mass. In this review we summarize recent findings on the contribution of remodeling to the pathological phenotype of asthma. We discuss how and why current treatment (such as corticosteroids) options fail to adequately treat remodeling. &copy; 2009 Weckmann et al, publisher and licensee Dove Medical Press Ltd.
Huynh, K., Oliver, B.G., Stelzer, S., Rawlinson, W. & Tovey, E. 2008, 'A new method for sampling and detection of exhaled respiratory virus aerosols', Clinical Infectious Diseases, vol. 46, no. 1, pp. 93-95.
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We have developed a mask sampler for exhaled respiratory viruses. Among a group of 9 patients with cold symptoms who had virus-positive nasal mucus specimens, as analyzed by multiplexed polymerase chain reaction, virus-positive mask samples were obtained
Oliver, B.G., Lim, S., Wark, P., Laza-Stanca, V., King, N., Black, J.L., Burgess, J., Roth, M. & Johnston, S.L. 2008, 'Rhinovirus exposure impairs immune responses to bacterial products in human alveolar macrophages', Thorax, vol. 63, no. 6, pp. 519-525.
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Background: Rhinovirus infection is responsible for considerable morbidity and mortality as the major cause of exacerbations of asthma, and is also known to induce exacerbations of cystic fibrosis and chronic obstructive pulmonary disease. Exacerbations of these diseases are also frequently associated with bacterial and atypical bacterial infection. Alveolar macrophages are the major immune cells in the airways and are important in defence against bacterial infections. Methods: The authors investigated whether rhinovirus modifies cytokine release, the pattern recognition receptor expression and phagocytosis by human alveolar macrophages in response to bacterial products. Results: Viable rhinovirus was detected in macrophages up to 3 days after exposure and viral RNA expression persisted for 10 days. Infectious but not UV inactivated rhinovirus increased tumour necrosis factor a (TNFa) and interleukin (IL)8 release by macrophages. In contrast, infectious rhinovirus impaired lipopolysaccharide and lipoteichoic acid induced TNFa and IL8 secretion by macrophages. Rhinovirus induced impairment of macrophage antibacterial immune responses did not involve IL10, prostaglandin E2 or downregulation of Toll-like receptor 2. Furthermore, the macrophage phagocytic response to labelled bacterial particles, but not to latex beads, was impaired. Conclusion: The authors have identified impairment of cytokine responses to bacterial lipopolysaccharide and lipoteichoic acid by alveolar macrophages in response to infectious rhinovirus. Virus induced impairment of antibacterial host defence has important implications in the pathogenesis of exacerbations of respiratory diseases.
Matsumoto, H., Moir, L., Oliver, B.G., Burgess, J., Roth, M., Black, J. & Mcparland, B. 2007, 'Comparison of gel contraction mediated by airway smooth muscle cells from patients with and without asthma', Thorax, vol. 62, no. 10, pp. 848-854.
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Backgrounds: Exaggerated bronchial constriction is the most significant and life threatening response of patients with asthma to inhaled stimuli. However, few studies have investigated the contractility of airway smooth muscle (ASM) from these patients.
Ammit, A.J., Moir, L.M., Oliver, B.G., Hughes, J.M., Alkhouri, H., Ge, Q., Burgess, J.K., Black, J.L. & Roth, M. 2007, 'Effect of IL-6 trans-signaling on the pro-remodeling phenotype of airway smooth muscle.', American journal of physiology. Lung cellular and molecular physiology, vol. 292, no. 1, pp. L199-L206.
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Increased levels of IL-6 are documented in asthma, but its contribution to the pathology is unknown. Asthma is characterized by airway wall thickening due to increased extracellular matrix deposition, inflammation, angiogenesis, and airway smooth muscle (ASM) mass. IL-6 binds to a specific membrane-bound receptor, IL-6 receptor-alpha (mIL-6Ralpha), and subsequently to the signaling protein gp130. Alternatively, IL-6 can bind to soluble IL-6 recpetor-alpha (sIL-6Ralpha) to stimulate membrane receptor-deficient cells, a process called trans-signaling. We discovered that primary human ASM cells do not express mIL-6Ralpha and, therefore, investigated the effect of IL-6 trans-signaling on the pro-remodeling phenotype of ASM. ASM required sIL-6Ralpha to activate signal transducer and activator 3, with no differences observed between cells from asthmatic subjects compared with controls. Further analysis revealed that IL-6 alone or with sIL-6Ralpha did not induce release of matrix-stimulating factors (including connective tissue growth factor, fibronectin, or integrins) and had no effect on mast cell adhesion to ASM or ASM proliferation. However, in the presence of sIL-6Ralpha, IL-6 increased eotaxin and VEGF release and may thereby contribute to local inflammation and vessel expansion in airway walls of asthmatic subjects. As levels of sIL-6Ralpha are increased in asthma, this demonstration of IL-6 trans-signaling in ASM has relevance to the development of airway remodeling.
Burgess, J., Oliver, B.G., Poniris, M., Ge, Q., Boustany, S., Cox, N., Moir, L., Johnson, P. & Black, J. 2006, 'A phosphodiesterase 4 inhibitor inhibits matrix protein deposition in airways in vitro', Journal of Allergy and Clinical Immunology, vol. 118, no. 3, pp. 649-657.
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Background: Airway smooth muscle (ASM) cells may contribute to airway remodeling through the release of growth factors, cytokines, and extracellular matrix (ECM) proteins. The effect of current asthma therapies on this release is not known. Objective: We
Oliver, B.G., Johnston, S., Baraket, M., Burgess, J., King, N., Roth, M., Lim, S. & Black, J. 2006, 'Increased proinflammatory responses from asthmatic human airway smooth muscle cells in response to rhinovirus infection', Respiratory Research, vol. 7, pp. 1-11.
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Background: Exacerbations of asthma are associated with viral respiratory tract infections, of which rhinoviruses (RV) are the predominant virus type. Airway smooth muscle is important in asthma pathogenesis, however little is known about the potential i
Oliver, B.G. & Black, J.L. 2006, 'Airway smooth muscle and asthma.', Allergology international : official journal of the Japanese Society of Allergology, vol. 55, no. 3, pp. 215-223.
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The airway smooth muscle is the key determinant of airway narrowing in asthma but its function in the absence of disease is unknown. Evidence for an intrinsic abnormality in the muscle in asthma is only just emerging. The airway smooth muscle is not merely a contractile cell, but also one which determines the composition of, and interacts with the extracellular matrix, and which may participate in inflammatory and allergic reactions and viral infections. The reason for the differences which have been observed in the in vitro properties of airway smooth muscle derived from asthmatic individuals may result from an inherent "supercontractility", an increased tendency to proliferate due to the absence of an inhibitory transcription factor C/EBP-alpha, the influence of an altered extracellular matrix and/or a decrease in release of factors such as PGE(2) which would under normal circumstances inhibit both proliferation and contraction. Although long acting beta agonists and corticosteroids are successful treatments for inflammation and bronchoconstriction, the structural changes which constitute airway remodelling may require additional therapeutic intervention, the nature of which will be determined by thorough investigation of the mechanisms underlying the asthmatic phenotype.
Sharma, R., von Haehling, S., Rauchhaus, M., Bolger, A.P., Genth-Zotz, S., Doehner, W., Oliver, B., Poole-Wilson, P.A., Volk, H.-.D., Coats, A.J.S., Adcock, I.M. & Anker, S.D. 2005, 'Whole blood endotoxin responsiveness in patients with chronic heart failure: the importance of serum lipoproteins.', European journal of heart failure, vol. 7, no. 4, pp. 479-484.
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BACKGROUND: Endotoxin [lipopolysaccharide (LPS)] may be an important stimulus for cytokine release in patients with chronic heart failure (CHF). We sought to investigate the relationship between whole blood endotoxin responsiveness and serum lipoprotein concentrations. It is not known if low-dose LPS is sufficient to stimulate immune activation. METHODS AND RESULTS: Whole blood from 32 CHF patients (mean age 66+/-2 years, NYHA class 2.7+/-0.2, five female) and 11 healthy control subjects (mean age 47+/-4 years, six female) was stimulated with LPS at nine different concentrations (0.001 to 10 ng/mL), and tumor necrosis factor (TNF-alpha) release was quantified. Reference standard endotoxin at concentrations of 0, 0.6, 1, and 3 EU/ml was added to whole blood from nine CHF patients (age 64+/-9.1 years, all NYHA class II, eight male) and incubated for 6 h, the TNF-alpha production being measured. Serum lipoproteins were quantified using standard techniques. In CHF patients, there was an inverse relationship between whole blood TNF-alpha release and serum cholesterol which was strongest at 0.6 ng/mL of LPS (r=-0.53, p=0.002). A similar although weaker relationship was found for serum HDL. No such correlation was found in healthy subjects or with serum LDL (all r(2)<0.1). Low concentrations of LPS induced a stepwise increase in TNF-alpha release from whole blood to concentrations well above those seen in CHF. CONCLUSIONS: Serum lipoproteins may play an important role in regulating LPS bioactivity in CHF. Very low LPS activity, at levels seen in vivo in CHF, can induce significant TNF-alpha production ex vivo.
Black, J.L., Ge, Q., Boustany, S., Johnson, P.R.A., Poniris, M.H., Glanville, A.R., Oliver, B.G.G., Moir, L.M. & Burgess, J.K. 2005, 'In vitro studies of lymphangioleiomyomatosis.', The European respiratory journal, vol. 26, no. 4, pp. 569-576.
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Lymphangioleiomyomatosis (LAM) is associated with abnormal airway smooth muscle that leads to the characteristic pathology of lung nodule formation and destruction of lung tissue. The current authors have previously identified abnormal behaviour of airway smooth muscle cells from patients with asthma. In this study, cells and tissue sections derived from patients with LAM (n=7), asthma (n=8), and nonasthmatic controls (n=9) were compared. The presence of the antigen human melanosome (HM)B-45 was investigated, along with the proliferation and release of extracellular matrix proteins, release of endogenous prostaglandin E2 (PGE2), vascular endothelial growth factor and connective tissue growth factor, and the expression of integrins. Positive HMB-45 staining was found in all LAM patients and no controls. Proliferation of LAM cells was not different from control cells nor was its inhibition by beta-agonists, corticosteroids, rapamycin or PGE2. However, endogenous PGE2 levels were markedly decreased in LAM cells, and this was associated with decreased expression of the inducible form of cyclooxygenase (COX-2). The increased levels of connective tissue growth factor seen in asthma cells were not observed in LAM. Elastin mRNA in response to transforming growth factor-beta stimulation was markedly lower in LAM cells than either asthma or control cells. In conclusion, lymphangioleiomyomatosis cells exhibit abnormal properties in vitro that may contribute to pathophysiology and symptomatology in patients with lymphangioleiomyomatosis.
Huntriss, J., Hinkins, M., Oliver, B., Harris, S.E., Beazley, J.C., Rutherford, A.J., Gosden, R.G., Lanzendorf, S.E. & Picton, H.M. 2004, 'Expression of mRNAs for DNA methyltransferases and methyl-CpG-binding proteins in the human female germ line, preimplantation embryos, and embryonic stem cells.', Molecular reproduction and development, vol. 67, no. 3, pp. 323-336.
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Recent evidence indicates that mammalian gametogenesis and preimplantation development may be adversely affected by both assisted reproductive and stem cell technologies. Thus, a better understanding of the developmental regulation of the underlying epigenetic processes that include DNA methylation is required. We have, therefore, monitored the expression, by PCR, of the mRNAs of DNA methyltransferases (DNMTs), methyl-CpG-binding domain proteins (MBDs), and CpG binding protein (CGBP) in a developmental series of amplified cDNA samples derived from staged human ovarian follicles, oocytes, preimplantation embryos, human embryonic stem (hES) cells and in similar murine cDNA samples. Transcripts of these genes were detected in human ovarian follicles (DNMT3A, DNMT3b1, DNMT3b4, DNMT1, MDBs1-4, MeCP2, CGBP), germinal vesicle (GV) oocytes (DNMT3A, DNMT3b1, DNMT1, MDBs1-4, MeCP2, CGBP), mature oocytes (DNMT3A, DNMT3b1, DNMT1, CGBP), and preimplantation embryos (DNMT3A, DNMT3b1, DNMT1, DNMT3L, MBD2, MDB4, CGBP). Differential expression of DNMT3B gene transcripts in undifferentiated (DNMT3b1) and in vitro differentiated human ES cells (DNMT3b3) further demonstrated an association of the DNMT3b1 transcript variant with totipotent and pluripotent human cells. Significantly, whilst the murine Dnmt3L gene is both expressed and essential for imprint establishment during murine oogenesis, transcripts of the human DNMT3L gene were only detected after fertilisation. Therefore, the mechanisms and/or the timing of imprint establishment may differ in humans.
Roche, N., Stirling, R.G., Lim, S., Oliver, B.G. & Chung, K.F. 2003, 'Regulation of protease-activated receptor-1 in mononuclear cells by neutrophil proteases.', Respiratory medicine, vol. 97, no. 3, pp. 228-233.
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Neutrophils and mononuclear cells are implicated in the pathogenesis of several inflammatory conditions including chronic obstructive pulmonary disease (COPD). Neutrophil-derived serine proteases, such as cathepsin G (CG) and neutrophil elastase (NE), may interact with mononuclear cells via protease-activated receptors (PARs) which are seven-transmembrane G protein-coupled receptors activated by proteolytic cleavage of the extracellular N-terminus, and which, on activation, induce the release of several mediators and cytokines. We determined whether CG and NE could affect PAR-1 expression and function in mononuclear cells. Human blood mononuclear cells were isolated from 20 healthy donors. Surface and intracellular receptor expression and calcium mobilisation (using the calcium chelator, FLUO3-AM) were studied by fluorescence-assisted cell sorting (FACS analysis). Positive controls, i.e. thrombin (0.1-100 mU/ml) and the PAR-1-activating peptide SFLLRN (100 microM) induced a rapid and transient intemalisation of PAR-1 in monocytes and lymphocytes. CG but not NE had a similar effect. By contrast, in monocytes intracellular calcium mobilisation was induced by thrombin and SFLLRN but not by CG and NE. Thus, CG can induce intracellular PAR-1 sequestration without activation of the receptor, and may act as an antagonist and prevent subsequent activation of PAR-1 in mononuclear cells. These findings may be of relevance to the pathogenesis of COPD.
Roche, N., Stirling, R.G., Lim, S., Oliver, B.G., Oates, T., Jazrawi, E., Caramori, G. & Chung, K.F. 2003, 'Effect of acute and chronic inflammatory stimuli on expression of protease-activated receptors 1 and 2 in alveolar macrophages.', The Journal of allergy and clinical immunology, vol. 111, no. 2, pp. 367-373.
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BACKGROUND: Protease-activated receptors 1 and 2 (PAR-1 and PAR-2) are 7-transmembrane G protein-coupled receptors activated by serine proteases in many cell types, including monocytes-macrophages, leading to the production of pro-inflammatory mediators, cytokines, and growth factors. OBJECTIVE: We determined the influence of chronic smoking and asthma on the expression of PAR-1 and PAR-2 receptors on alveolar macrophages (AMs). METHODS: We used RT-PCR and immunocytochemistry with confocal microscopy to determine mRNA and protein expression of PAR-1 and PAR-2 in AMs obtained from healthy smokers, asthmatic patients, and healthy subjects. In addition, we examined the effect of IL-1beta and LPS. RESULTS: PAR1 mRNA was decreased, whereas PAR2 mRNA was increased in 24-hour cultured AMs from smokers when compared with values in AMs from healthy subjects. Paradoxically, there was a higher degree of PAR-1 protein staining in AMs from smokers, whereas PAR-2 staining was similar in smokers and healthy subjects. PAR-1 and PAR-2 mRNA and protein expression were similar in asthmatic patients and control subjects. IL-1beta and LPS had no effect on PAR1 and PAR2 gene expression by AMs. CONCLUSIONS: There is a dissociation between gene and protein expression of PAR-1 and PAR-2. PAR-1 protein overexpression in AMs from smokers might be important in the pathophysiology of chronic airways disease.
Huntriss, J., Gosden, R., Hinkins, M., Oliver, B., Miller, D., Rutherford, A.J. & Picton, H.M. 2002, 'Isolation, characterization and expression of the human Factor In the Germline alpha (FIGLA) gene in ovarian follicles and oocytes.', Molecular human reproduction, vol. 8, no. 12, pp. 1087-1095.
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The Factor In the Germline alpha (FIGalpha) transcription factor regulates expression of the zona pellucida proteins ZP1, ZP2 and ZP3 and is essential for folliculogenesis in the mouse. Using the published mouse Figla sequence, BLAST searches identified a human chromosome 2 BAC clone with high sequence identity. Using PCR primers derived from this clone, amplicons derived from ovarian follicles and mature oocytes revealed 100% identity with the appropriate human BAC clone, the expected homology with the mouse Figla gene sequence, and homology on translation with the FIGalpha protein identified in the Japanese rice fish, medaka (Oryzias latipes). PCR expression profiling of this transcript revealed FIGLA mRNA expression in cDNA derived from ovarian follicles (5/5 samples from the primordial through to the secondary stage) mature oocytes (6/9 samples), and less frequently in preimplantation embryos (2/7 samples). Subsequent BLAST searches revealed the predicted full length coding sequence of the human FIGalpha protein which demonstrates 68 and 25% similarity overall to mouse and medaka proteins respectively, with 96 and 57% identity respectively within the basic helix-loop-helix region. This confirms our identification of the human homologue for this gene which maps to chromosome 2p12. Further work is required to understand its role in normal human oocyte development and the potential involvement in human infertility.
Oliver, B., Tomita, K., Keller, A., Caramori, G., Adcock, I., Chung, K.F., Barnes, P.J. & Lim, S. 2001, 'Low-dose theophylline does not exert its anti-inflammatory effects in mild asthma through upregulation of interleukin-10 in alveolar macrophages.', Allergy, vol. 56, no. 11, pp. 1087-1090.
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BACKGROUND: There is accumulating evidence that theophylline has anti-inflammatory or immunomodulatory effects. This may be, in part, mediated via an upregulation in the production of the anti-inflammatory cytokine interleukin (IL)-10. We determined whether low-dose theophylline (LDT) would increase the production of IL-10, and attenuate the production of proinflammatory cytokines by alveolar macrophages. METHODS: In a double-blind, placebo-controlled, crossover study involving 15 steroid-free patients with mild asthma, fiberoptic bronchoscopy and bronchoalveolar lavage (BAL) were performed at the end of the treatment and placebo periods. Alveolar macrophages were cultured in vitro, and we measured their release of IL-10, GM-CSF, and TNF-alpha. We also measured IL-10 production in whole blood together with the number of monocytes and T cells expressing intracellular IL-10 by flow cytometry. RESULTS: LDT did not increase the production of IL-10, or attenuate the production of GM-CSF or TNF-alpha by alveolar macrophages. However, after theophylline treatment, there was a significant reduction in mean (SD) (95% CI) BAL eosinophil number from 3.4 (1.7)% (95% CI 2.4-4.4) to 1.7 (1.0)% (95% CI 1.1-2.3) compared with placebo (P<0.05). Similarly, there was no increase in whole-blood IL-10 release or in the number of monocytes and T cells expressing intracellular IL-10 after treatment. CONCLUSIONS: LDT has an anti-inflammatory effect in asthma; however, this effect is not mediated via the production of IL-10 or the attenuation of GM-CSF or TNF-alpha. The mechanisms of theophylline activity remain to be determined.
Lim, S., Tomita, K., Caramori, G., Jatakanon, A., Oliver, B., Keller, A., Adcock, I., Chung, K.F. & Barnes, P.J. 2001, 'Low-dose theophylline reduces eosinophilic inflammation but not exhaled nitric oxide in mild asthma.', American journal of respiratory and critical care medicine, vol. 164, no. 2, pp. 273-276.
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Theophylline is well-established in the management of asthma, and there is some evidence of an antiinflammatory effect in asthma. It is not known whether theophylline affects inflammatory markers such as sputum eosinophils and exhaled nitric oxide (NO) in patients with mild asthma not receiving inhaled steroid therapy. In a double-blind, placebo-controlled, cross-over study of 15 patients with mild asthma, we assessed the effect of low-dose theophylline therapy (250 mg twice per day) on eosinophils in induced sputum, bronchoalveolar lavage (BAL) and airway biopsies at the end of both the treatment and placebo periods. Measurements of exhaled nitric oxide (NO) were made at the end of the active and placebo treatment periods of 5 wk each. Low-dose theophylline (mean serum level, 6.1 mg/L) led to a significant reduction in mean (95% confidence interval [CI]) sputum eosinophils from 11.3% (7.80-14.76%) to 8.0% (5.46-10.44%), BAL eosinophils from 3.4% (2.4-4.4%) to 1.7% (1.1-2.3%) and biopsy eosinophils from 1.83% (0.76-2.89%) to 1.20% (0.27-2.13%) compared with placebo (all p < 0.05). There was no significant change in levels of exhaled NO or improvement in lung function and bronchial responsiveness. Low-dose theophylline induced antiinflammatory effects in asthma, reflected by a fall in airway eosinophils with no change in exhaled NO or changes in lung function.
Lim, S., Roche, N., Oliver, B.G., Mattos, W., Barnes, P.J. & Chung, K.F. 2000, 'Balance of matrix metalloprotease-9 and tissue inhibitor of metalloprotease-1 from alveolar macrophages in cigarette smokers. Regulation by interleukin-10.', American journal of respiratory and critical care medicine, vol. 162, no. 4 Pt 1, pp. 1355-1360.
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An imbalance between proteases and antiproteases may play a role in emphysema, which is characterized by increased degradation of extracellular matrix, and in airway remodeling in chronic bronchitis and asthma, in which there is increased collagen deposition. We assessed the effect of smoking on release of matrix metalloprotease-9 (MMP-9) and of its inhibitor, tissue inhibitor of metalloprotease-1 (TIMP-1), from alveolar macrophages, and determined the effects of proinflammatory (interleukin [IL]-1beta and lipopolysaccharide [LPS]) and antiinflammatory (IL-10) stimuli on the release of MMP-9 and TIMP-1. We performed bronchoalveolar lavage in 11 smokers and 11 nonsmokers, and cultured airway macrophages in the presence of control medium, IL-1beta, and LPS. Airway macrophages from smokers released greater amounts of MMP-9 and TIMP-1 at baseline and in response to IL-1beta and LPS than did those of nonsmokers. Airway macrophages from smokers produced more TNF-alpha and IL-10. IL-10 increased TIMP-1 release without modifying that of MMP-9, leading to a decrease in the MMP-9 to TIMP-1 ratio. Anti-IL-10 antibody had no effect on MMP-9 production induced by LPS. We conclude that the release of proteases and antiproteases by airway macrophages is increased in cigarette smokers, and can be regulated by exogenous IL-10.
Aaron, J.E., Oliver, B., Clarke, N. & Carter, D.H. 1999, 'Calcified microspheres as biological entities and their isolation from bone.', The Histochemical journal, vol. 31, no. 7, pp. 455-470.
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Calcified microspheres, about 1 microm in diameter, appear at sites of bone formation where they invest the collagenous matrix, become confluent and disappear. Evidence that the particle boundaries are not lost with compaction but merely deformed is supported in section by the granular histochemical staining of the inorganic phase for bone salt, lipid, fibronectin and acid phosphatase in osteomalacic, acid-etched and normal human bone. Their persistence as discrete objects is confirmed by the application of methods for their isolation from the collagenous matrix of immature mouse calvarium and mature bovine femur. Five methods have been used to extract them and include (i) biochemical, (ii) chemical, (iii) mechanical, (iv) pyrogenous and (v) biological separation. Under the optical microscope, all isolates consisted of similar discrete objects and bridged assemblies, whose birefringence varied with treatment. After decalcification, their organic 'ghosts' remained. Each isolated microsphere had a complex substructure of clusters of non-collagenous calcified filaments surrounding a less dense centre. The filaments were 5 nm in diameter with a 5 nm periodicity and regular fine interfilamentous connections. It is concluded that the microspheres are independent, complex, pervasive and central to the containment (i.e. packaging) of calcium phosphate in bone. Their extraction will enable further analysis.

Other

Sharma, P., Yi, R., Nayak, A., Wang, N., Tang, F., Knight, M., Pan, S., Oliver, B. & Deshpande, D. 2017, 'Bitter Taste Receptor Agonists Mitigate Features of Allergic Asthma in Mice'.
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Asthma is characterized by airway inflammation, mucus secretion, remodeling and hyperresponsiveness (AHR). Recent research has established the bronchodilatory effect of bitter taste receptor (TAS2R) agonists in various models. Comprehensive pre-clinical studies aimed at establishing effectiveness of TAS2R agonists in disease models are lacking. Here we aimed to determine the effect of TAS2R agonists on features of asthma. Further, we elucidated a mechanism by which TAS2R agonists mitigate features of asthma. Asthma was induced in mice using intranasal house dust mite or aerosol ova-albumin challenge, and chloroquine or quinine were tested in both prophylactic and treatment models. Allergen challenge resulted in airway inflammation as evidenced by increased immune cells infiltration and release of cytokines and chemokines in the lungs, which were significantly attenuated in TAS2R agonists treated mice. TAS2R agonists attenuated features of airway remodeling including smooth muscle mass, extracellular matrix deposition and pro-fibrotic signaling, and also prevented mucus accumulation and development of AHR in mice. Mechanistic studies using human neutrophils demonstrated that inhibition of immune cell chemotaxis is a key mechanism by which TAS2R agonists blocked allergic airway inflammation and exerted anti-asthma effects. Our comprehensive studies establish the effectiveness of TAS2R agonists in mitigating multiple features of allergic asthma.

I have a growing international reputation and have numerous active collaborations with researchers in Australia, Europe, Asia and the USA.

My active academic collaborations external to UTS  include:

Professor Philip Hansbro (Uni Newcastle)

Professor Ian Addcock (imperial Collage, UK)

Dr Corry-Anke Brandsma  (Uni Groningen, the Nethterlands)

Professor Dirkje Postma and Win Timens (Uni Groningen, the Nethterlands)

Professor Patricia Finn (Uni Chicgo)

I also have a number of active collaborations with Industry, including GSK, AZ, and Pharmaxsis.