Publications
91 results found
Wilson M, Petrie J, Shaw M, et al., 2016, High Fat Feeding Protects Mice From Ventilator-Induced Lung Injury Via A Neutrophil-Independent Mechanism, International Conference of the American-Thoracic-Society (ATS), Publisher: AMER THORACIC SOC, ISSN: 1073-449X
Soni S, Yoshida M, Woods S, et al., 2015, Microvesicles derived from alveolar macrophages mediate epithelial activation via a TNF dependant mechanism, Publisher: EUROPEAN RESPIRATORY SOC JOURNALS LTD, ISSN: 0903-1936
Patel BV, Tatham KC, Wilson MR, et al., 2015, In vivo compartmental analysis of leukocytes in mouse lungs, American Journal of Physiology-Lung Cellular and Molecular Physiology, Vol: 309, Pages: L639-L652, ISSN: 1522-1504
The lung has a unique structure consisting of three functionally different compartments (alveolar, interstitial, and vascular) situated in an extreme proximity. Current methods to localize lung leukocytes using bronchoalveolar lavage and/or lung perfusion have significant limitations for determination of location and phenotype of leukocytes. Here we present a novel method using in vivo antibody labelling to enable accurate compartmental localization/quantification and phenotyping of mouse lung leukocytes. Anesthetized C57BL/6 mice received combined in vivo intravenous and intratracheal labelling with fluorophore-conjugated anti-CD45 antibodies, and lung single cell suspensions were analyzed by flow cytometry. The combined in vivo intravenous and intratracheal CD45 labelling enabled robust separation of the alveolar, interstitial, and vascular compartments of the lung. In naive mice, the alveolar compartment consisted predominantly of resident alveolar macrophages. The interstitial compartment, gated by events negative for both intratracheal and intravenous CD45 staining, showed two conventional dendritic cell populations, as well as a Ly6C(lo) monocyte population. Expression levels of MHCII on these interstitial monocytes were much higher than the vascular Ly6C(lo) monocyte populations. In mice exposed to acid-aspiration induced lung injury, this protocol also clearly distinguished the three lung compartments showing the dynamic trafficking of neutrophils and exudative monocytes across the lung compartments during inflammation and resolution. This simple in vivo dual labelling technique substantially increases the accuracy and depth of lung flow cytometric analysis, facilitates a more comprehensive examination of lung leukocyte pools, and enables the investigation of previously poorly defined 'interstitial' leukocyte populations during models of inflammatory lung diseases.
Patel BV, Tatham KC, Wilson MR, et al., 2015, In Vivo Compartmental Labeling of the Mouse Lung, American Thoracic Society, Publisher: ATS Journals, Pages: A3926-A3926, ISSN: 1073-449X
ShareModerators: G.N. Maksym, PhD, R.S. Harris, MD, J.C. Sieren, PhDSession Info: Mini Symposium, [C18] STATE OF PLAY IN RESPIRATORY STRUCTURE AND FUNCTION: SEEING IS BELIEVINGDay/Date: Tuesday, May 19, 2015Session Time: 9:30 AM - 11:30 AMRoom: Mile High Ballroom 2C/3C (Lower Level)Location: Colorado Convention CenterIn Vivo Compartmental Labeling of the Mouse Lung, [Publication Page: A3926]B.V. Patel, MBBS MRCP FRCA PhD, K.C. Tatham, MBBS, M.R. Wilson, PhD, K.P. O'Dea, PhD, M. Takata, MDLondon/UKRationaleCurrent methods for compartmental analyses of lungs using bronchoalveolar lavage (BAL) and/or lung perfusion have significant limitations for accurate determination of location and phenotype of lung leukocytes. BAL retrieves only <25% of intra-alveolar cells (1) and lung perfusion fails to remove vascular marginated cell populations (2). We present a novel method of in vivo antibody labelling to facilitate the accurate compartmental investigation of mouse lung leukocytes by flow cytometry.MethodsAnesthetized C57BL6 mice underwent tracheostomy and venous cannulation. An anti-CD45 antibody (PE-conjugated) was injected intravenously and allowed to circulate for 5 minutes. Mice were then exsanguinated and lungs immediately flooded intratracheally with another anti-CD45 antibody (PE-Cy5-conjugated). Lungs were harvested, and single cell suspensions prepared for flow cytometric analysis using a panel of myeloid markers. This in vivo labelling protocol was also applied to mice exposed to acid-aspiration induced lung injury (3). To confirm the separation between the vascular and interstitial cell populations, a group of mice underwent intravenous labelling alone followed by lung perfusion for 15 minutes using an isolated-perfused lung apparatus (2).ResultsThe combined in vivo intravenous and intratracheal CD45 labelling enabled robust separation of the alveolar, interstitial, and vascular compartments of the lung by flow cytometry, with <0.3% of leukocytes staining
Woods SJ, Waite AAC, O'Dea KP, et al., 2015, Kinetic profiling of in vivo lung cellular inflammatory responses to mechanical ventilation, AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, Vol: 308, Pages: L912-L921, ISSN: 1040-0605
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- Citations: 33
Lax S, Wilson MR, Takata M, et al., 2014, Using a non-invasive assessment of lung injury in a murine model of acute lung injury, BMJ Open Respiratory Research, Vol: 1, ISSN: 2052-4439
Arterial oxygen saturation has not been assessed sequentially in conscious mice as a direct consequence of an in vivo murine model of acute lung injury. Here, we report daily changes in arterial oxygen saturation and other cardiopulmonary parameters by using infrared pulse oximetry following intratracheal lipopolysaccharide (IT-LPS) for up to 9 days, and following IT-phosphate buffered saline up to 72 h as a control. We show that arterial oxygen saturation decreases, with maximal decline at 96 h post IT-LPS. Blood oxygen levels negatively correlate with 7 of 10 quantitative markers of murine lung injury, including neutrophilia and interleukin-6 expression. This identifies infrared pulse oximetry as a method to non-invasively monitor arterial oxygen saturation following direct LPS instillations.
Wakabayashi K, Wilson MR, Tatham KC, et al., 2014, Volutrauma, but not Atelectrauma, Induces Systemic Cytokine Production by Lung-Marginated Monocytes, Critical Care Medicine, Vol: 42, Pages: e49-e57, ISSN: 0090-3493
Objectives: Ventilator-induced lung injury has substantive impact on mortality of patients with acute respiratory distress syndrome. Although low tidal volume ventilation has been shown to reduce mortality, clinical benefits of open-lung strategy are controversial. In this study, we investigated the impact of two distinct forms of ventilator-induced lung injury, i.e., volutrauma and atelectrauma, on the progression of lung injury and inflammation, in particular alveolar and systemic cytokine production.Design: Ex vivo study.Setting: University research laboratory.Subjects: C57BL/6 mice.Interventions: Isolated, buffer-perfused lungs were allocated to one of three ventilatory protocols for 3 hours: control group received low tidal volume (7 mL/kg) with positive end-expiratory pressure (5 cm H2O) and regular sustained inflation; high-stretch group received high tidal volume (30–32 mL/kg) with positive end-expiratory pressure (3 cm H2O) and sustained inflation; and atelectasis group received the same tidal volume as control but neither positive end-expiratory pressure nor sustained inflation.Measurements and Main Results: Both injurious ventilatory protocols developed comparable levels of physiological injury and pulmonary edema, measured by respiratory system mechanics and lavage fluid protein. High-stretch induced marked increases in proinflammatory cytokines in perfusate and lung lavage fluid, compared to control. In contrast, atelectasis had no effect on perfusate cytokines compared to control but did induce some up-regulation of lavage cytokines. Depletion of monocytes marginated within the lung microvasculature, achieved by pretreating mice with IV liposome-encapsulated clodronate, significantly attenuated perfusate cytokine levels, especially tumor necrosis factor, in the high-stretch, but not atelectasis group.Conclusions: Volutrauma (high-stretch), but not atelectrauma (atelectasis), directly activates monocytes within the pulm
Soni S, Wilson MR, O'Dea K, et al., 2014, Microvesicles Are Sequentially Released From Different Intra-Alveolar Cells In A Mouse Model Of Acute Lung Injury, Publisher: AMER THORACIC SOC, ISSN: 1073-449X
Wakabayashi K, Wilson M, Tatham K, et al., 2013, High-stretch, but not atelectasis, causes systemic cytokine release by lung-marginated monocytes, Publisher: EUROPEAN RESPIRATORY SOC JOURNALS LTD, ISSN: 0903-1936
Lax S, Wilson M, Takata M, et al., 2013, The role of pre-receptor glucocorticoid metabolism in regulating the severity and persistence of murine lung injury, Publisher: EUROPEAN RESPIRATORY SOC JOURNALS LTD, ISSN: 0903-1936
Patel BV, Wilson MR, O'Dea KP, et al., 2013, TNF-Induced Death Signaling Triggers Alveolar Epithelial Dysfunction in Acute Lung Injury, J Immunol, Vol: 190, Pages: 4274-4282
Wilson MR, Takata M, 2013, Inflammatory mechanisms of ventilator-induced lung injury: a time to stop and think?, Anaesthesia, Vol: 68, Pages: 175-178
Wakabayashi K, Wilson MR, Patel BV, et al., 2013, Inhibition Of TNF Receptor p55 By A Domain Antibody Attenuates Acid-Induced Lung Injury In Mice, American Thoracic Society
RationaleTumor necrosis factor (TNF) is strongly implicated in the evolution of acute lung injury (ALI), but its potential as a therapeutic target has been hampered by its complex biology. TNF signals through two cell surface receptors, p55 and p75, which play differential roles in ALI, with p55 promoting injury while p75 opposes this. We have recently shown that selective p55 blockade using novel domain antibody technology attenuates ventilator-induced lung injury (VILI) in mice. To further translate these findings to the bedside, we explored the potential for selective blockade of p55 in a clinically relevant mouse model of acid-induced ALI.MethodsMale C57BL6 mice were pretreated with intranasal administration of either specific domain antibody to p55 (dAb) or non-specific control domain antibody (‘dummy’). Four hours later, the mice were either instrumented for analysis without further instillation (i.e. baseline) or intratracheally challenged with hydrochloric acid (0.1M, 75μl) via an oro-tracheal tube to induce ALI. All acid-challenged animals were instrumented for analysis at 24 hours after acid instillation. Analysis consisted of mice being anesthetized, tracheostomized, and mechanically ventilated in order to evaluate PaO2/FiO2 ratio and respiratory mechanics. The left lung was taken for determining wet/dry ratio and a lunglavage sample was taken from the right lung to measure protein levels and neutrophil counts.ResultsHydrochloric acid instillation in the dummy group resulted in severe ALI, represented by hypoxemia, elastance increase, pulmonary edema and inflammation (Table 1). These mice were also visually very sick, showing considerably reduced locomotion and responsiveness. In contrast, pretreatment with p55 dAb dramatically improved the clinical symptoms, along with significant attenuation in all the physiological markers of lung injury. However, the dAb treatment did not significantly attenuate lung neutrophil recruitment represented b
Lax S, Wilson MR, Takata M, et al., 2012, THE ROLE OF PRE-RECEPTOR GLUCOCORTICOID METABOLISM IN REGULATING THE SEVERITY AND PERSISTENCE OF MURINE LUNG INJURY, Winter Meeting of the British-Thoracic-Society 2012, Publisher: BMJ PUBLISHING GROUP, Pages: A28-A28, ISSN: 0040-6376
Wilson MR, Patel BV, Takata M, 2012, Ventilation with "clinically relevant" high tidal volumes does not promote stretch-induced injury in the lungs of healthy mice, CRITICAL CARE MEDICINE, Vol: 40, Pages: 2850-2857, ISSN: 0090-3493
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- Citations: 49
Petrie J, Wilson MR, Patel BV, et al., 2012, Obesity induced by high fat feeding attenuates ventilator-induced lung injury in mice., European Respiratory Society Annual Congress
BackgroundRetrospective analysis of Intensive Care data suggests that obesity may confer a survival advantage in Acute Lung Injury (ALI). Development of ventilator-induced lung injury (VILI) is a major determinant of ALI mortality. We have therefore investigated the impact of high fat diet-induced obesity on VILI in mice.MethodsMale C57BL/6 mice were fed high fat diet for a minimum of 13 weeks, resulting in a mean body weight 30% greater than lean age-matched controls. Animals were anaesthetised and ventilated with high-stretch, standardised as plateau pressure (Pplat) 35-37cmH2O. Tidal volumes were similar between groups (mean ∽1080μl). Mice were ventilated for 180mins or until Pplat increased by 20%. Lung tissue was harvested for wet:dry ratio or processed to a single cell suspension for leukocyte quantification by flow cytometry.ResultsHigh stretch ventilation induced increases in Pplat and lung wet:dry ratio, and a decrease in paO2 in lean mice, which were all significantly attenuated in obese animals. Leukocyte recruitment (cells/g dry lung mass) also tended to be reduced. High fat feeding attenuates pulmonary oedema and lung dysfunction associated with VILI in mice. Numerous metabolic and immunological differences exist between lean and obese subjects. Exploring the mechanisms behind this obesity-mediated protection from VILI may lead to identification of novel pathways and therapeutic targets.Funded by: BJA/Royal College of Anaesthetists.
Patel BV, Wilson MR, Takata M, 2012, Resident alveolar macrophages mediate early alveolar epithelial death signaling and dysfunction., European Respiratory Society Annual Congress
Acute lung injury (ALI) is characterized by alveolar epithelial dysfunction. We previously showed that early epithelial dysfunction was specifically mediated through tumor necrosis factor (TNF) p55 receptor signaling [1]. This study examined the contribution of resident alveolar macrophages (AM) to this phenomenon following acid aspiration.C57Bl6 mice were treated intratracheally with liposomes containing either clodronate or PBS. After 48 hours, they underwent intratracheal instillation of hydrochloric acid followed by mechanical ventilation to assess respiratory parameters. Oxygenation, respiratory elastance, alveolar TNF concentration, lung caspase-8 activity and alveolar fluid clearance (AFC) were measured at 90 minutes after acid instillation.Clodronate liposomes produced an 80% depletion of AMs. AM depletion significantly improved the deterioration in respiratory elastance (cmH2O/µl: PBS=0.06±0.008; CLOD=0.05±0.004; p<0.05) and PaO2:FiO2 (PBS=304±113; CLOD=426±41; P<0.05) induced by acid instillation. Additionally, alveolar TNF was significantly reduced (pg/ml: PBS=46.5±25.8; CLOD=15.5±2.7; P<0.05), along with attenuated lung caspase-8 activity (arbitrary units: PBS=14763±5466; CLOD=7135±372; P<0.01), and improved AFC (%/30min: PBS=3.8±2.6; CLOD=7.1±2.4; P<0.05). Caspase-8 activity showed an inverse correlation to AFC (Pearson r=-0.766; P<0.0001) implying epithelial death receptor activation.These data suggests that during ALI induced by acid aspiration, epithelial dysfunction and hypoxemia are a result of epithelial cell death receptor activation by alveolar macrophage-derived TNF.[1] Patel et al. Intensive Care Med. 2011;37(Supplement):S205Supported by Wellcome Trust, UK.
Lax S, Wilson M, Takata M, et al., 2012, The role of pre-receptor glucocorticoid metabolism in regulating the severity of ALI, Publisher: EUROPEAN RESPIRATORY SOC JOURNALS LTD, ISSN: 0903-1936
Patel BV, Wilson MR, Takata M, 2012, Resolution of acute lung injury and inflammation – a translational mouse model., European Respiratory Journal, Vol: 39, Pages: 1162-1170
Bertok S, Wilson MR, Morley PJ, et al., 2012, Selective inhibition of intra-alveolar p55 TNF receptor attenuates ventilator-induced lung injury, Thorax, Vol: 67, Pages: 244-251
Wilson MR, Patel BV, Takata M, 2011, DO "CLINICALLY RELEVANT" TIDAL VOLUMES REALLY CAUSE VENTILATOR-INDUCED LUNG INJURY IN MICE?, Winter Meeting of the British-Thoracic-Society, Publisher: B M J PUBLISHING GROUP, Pages: A36-A37, ISSN: 0040-6376
O'Dea KP, Dokpesi JO, Tatham KC, et al., 2011, Regulation of monocyte subset proinflammatory responses within the lung microvasculature by the p38 MAPK/MK2 pathway, AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, Vol: 301, Pages: L812-L821, ISSN: 1040-0605
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- Citations: 29
Patel BV, Wilson MR, Takata M, 2011, The p55 TNF receptor promotes alveolar epithelial dysfunction in experimental lung injury., European Society of Intensive Care Medicine
Dorr AD, Wilson MR, Wakabayashi K, et al., 2011, Sources of alveolar soluble TNF receptors during acute lung injury of different etiologies, JOURNAL OF APPLIED PHYSIOLOGY, Vol: 111, Pages: 177-184, ISSN: 8750-7587
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- Citations: 14
Bertok S, Wilson MR, Dorr AD, et al., 2011, Characterization of TNF receptor subtype expression and signaling on pulmonary endothelial cells in mice, AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, Vol: 300, Pages: L781-L789, ISSN: 1040-0605
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- Citations: 22
Wakabayashi K, Wilson MR, O'Dea KP, et al., 2011, Atelectasis Induces Chemokine Upregulation And Lung Injury In The Isolated Perfused Mouse Lung, Publisher: AMER THORACIC SOC, ISSN: 1073-449X
Bertok S, Wilson MR, Morley PJ, et al., 2010, BLOCKADE OF INTRAALVEOLAR P55 TNF-RECEPTOR SIGNALLING BY A DOMAIN ANTIBODY DECREASES INFLAMMATION AND OEDEMA IN AN <i>IN VIVO</i> MOUSE MODEL OF VENTILATOR-INDUCED LUNG INJURY, British-Thoracic-Society-Winter-Meeting 2010, Publisher: B M J PUBLISHING GROUP, Pages: A1-A1, ISSN: 0040-6376
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- Citations: 1
Waite AC, O'Dea KP, Wilson MR, et al., 2010, FLOW CYTOMETRIC DETECTION OF INTRACELLULAR ACTIVATION MARKERS OF PULMONARY CELLS DURING ACUTE LUNG INJURY, Publisher: B M J PUBLISHING GROUP, Pages: A182-A183, ISSN: 0040-6376
Wakabayashi K, Wilson MR, O'Dea KP, et al., 2010, COMPARISON OF HIGH-STRETCH VERSUS ATELECTASIS IN THE PATHOPHYSIOLOGY OF VENTILATOR-INDUCED LUNG INJURY USING THE MOUSE ISOLATED PERFUSED LUNG, British-Thoracic-Society-Winter-Meeting 2010, Publisher: B M J PUBLISHING GROUP, Pages: A49-A50, ISSN: 0040-6376
Bertok S, Wilson MR, Morley PJ, et al., 2010, SPECIFIC INHIBITION OF INTRA-ALVEOLAR P55 TNF RECEPTOR SIGNALLING BY A DOMAIN ANTIBODY ATTENUATES VENTILATOR-INDUCED LUNG INJURY IN MICE, 23rd Annual Meeting of the European-Society-of-Intensive-Care-Medicine, Publisher: SPRINGER, Pages: S196-S196, ISSN: 0342-4642
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- Citations: 1
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