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W Tai,
L Roberts,
A Seryshev,
J M Gubatan,
C S Bland,
R Zabriskie,
S Kulkarni,
L Soong,
I Mbawuike,
B Gilbert, F Kheradmand,
D B Corry
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ABSTRACT: All commercial influenza vaccines elicit antibody responses that protect against seasonal infection, but this approach is limited by the need for annual vaccine reformulation that precludes efficient responses against epidemic and pandemic disease. In this study we describe a novel vaccination approach in which a nanoparticulate, liposome-based agent containing short, highly conserved influenza-derived peptides is delivered to the respiratory tract to elicit potent innate and selective T cell-based adaptive immune responses. Prepared without virus-specific peptides, mucosal immunostimulatory therapeutic (MIT) provided robust, but short-lived, protection against multiple, highly lethal strains of influenza in mice of diverse genetic backgrounds. MIT prepared with three highly conserved epitopes that elicited virus-specific memory T-cell responses but not neutralizing antibodies, termed MITpep, provided equivalent, but more durable, protection relative to MIT. Alveolar macrophages were more important than dendritic cells in determining the protective efficacy of MIT, which induced both canonical and non-canonical antiviral immune pathways. Through activation of airway mucosal innate and highly specific T-cell responses, MIT and MITpep represent novel approaches to antiviral protection that offer the possibility of universal protection against epidemic and pandemic influenza.
Mucosal Immunology 03/2011; 4(2):197-207. · 6.96 Impact Factor
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P Porter,
S C Susarla,
S Polikepahad,
Y Qian,
J Hampton,
A Kiss,
S Vaidya,
S Sur,
V Ongeri,
T Yang,
G L Delclos,
S Abramson, F Kheradmand,
D B Corry
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ABSTRACT: Active fungal proteinases are powerful allergens that induce experimental allergic lung disease strongly resembling atopic asthma, but the precise relationship between proteinases and asthma remains unknown. Here, we analyzed dust collected from the homes of asthmatic children for the presence and sources of active proteinases to further explore the relationship between active proteinases, atopy, and asthma. Active proteinases were present in all houses and many were derived from fungi, especially Aspergillus niger. Proteinase-active dust extracts were alone insufficient to initiate asthma-like disease in mice, but conidia of A. niger readily established a contained airway mucosal infection, allergic lung disease, and atopy to an innocuous bystander antigen. Proteinase produced by A. niger enhanced fungal clearance from lung and was required for robust allergic disease. Interleukin 13 (IL-13) and IL-5 were required for optimal clearance of lung fungal infection and eosinophils showed potent anti-fungal activity in vitro. Thus, asthma and atopy may both represent a protective response against contained airway infection due to ubiquitous proteinase-producing fungi.
Mucosal Immunology 09/2009; 2(6):504-17. · 6.96 Impact Factor
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ABSTRACT: We show that the interaction of the Yersinia surface protein, invasin, with rabbit synovial fibroblasts mediates bead phagocytosis and induces expression of interleukin 1alpha (IL-1alpha), tumor necrosis factor-alpha (TNF-alpha) and MMP-1/collagenase-1 (CL-1). Presentation of invasin as a ligand on the surface of 4.5 microm beads induced phagocytosis and increased CL-1 expression 20-fold after 24 hours. By contrast, presentation of invasin as a spreading substrate did not induce CL-1 expression. CL-1 induction following phagocytosis of invasin-coated beads was mediated by a mechanism dependent on high-affinity binding to beta1 integrins and the function of the small GTPase RhoA. Expression of a function-perturbing mutant, RhoAN19, abrogated bead-induced CL-1 expression. RhoA activation coupled bead phagocytosis with signal transduction because expression of constitutively active mutant RhoV14 was sufficient to trigger CL-1 expression. The signal-transduction cascade elicited by bead phagocytosis triggered NFkappaB activation, stimulating a proinflammatory cellular response with transient increases in TNF-alpha production that peaked at 2 hours and induction of IL-1alpha that was sustained for at least 10 hours. Inhibition of IL-1alpha function by blocking antibodies or IL-1 receptor antagonist showed that IL-1alpha is the autocrine intermediary for subsequent CL-1 induction.
Journal of Cell Science 10/2001; 114(Pt 18):3333-43. · 6.11 Impact Factor
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ABSTRACT: Immunoglobulin E (IgE) is believed to be one of the major mediators of immediate hypersensitivity reactions that underlie atopic conditions such as urticaria, seasonal allergy, asthma and anaphylaxis. Factors that control IgE production are therefore essential to the pathogenesis of these important afflictions. But a complete understanding of this topic is lacking, while new data have raised questions regarding the precise role of IgE in atopic disease. Evolving concepts of IgE production and elimination are likely to clarify the importance of IgE in health and disease.
Nature 12/1999; 402(6760 Suppl):B18-23. · 36.28 Impact Factor
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ABSTRACT: Integrin-mediated reorganization of cell shape leads to an altered cellular phenotype. Disruption of the actin cytoskeleton, initiated by binding of soluble antibody to alpha5beta1 integrin, led to increased expression of the collagenase-1 gene in rabbit synovial fibroblasts. Activation of the guanosine triphosphate-binding protein Rac1, which was downstream of the integrin, was necessary for this process, and expression of activated Rac1 was sufficient to increase expression of collagenase-1. Rac1 activation generated reactive oxygen species that were essential for nuclear factor kappa B-dependent transcriptional regulation of interleukin-1alpha, which, in an autocrine manner, induced collagenase-1 gene expression. Remodeling of the extracellular matrix and consequent alterations of integrin-mediated adhesion and cytoarchitecture are central to development, wound healing, inflammation, and malignant disease. The resulting activation of Rac1 may lead to altered gene regulation and alterations in cellular morphogenesis, migration, and invasion.
Science 05/1998; 280(5365):898-902. · 31.20 Impact Factor
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ABSTRACT: Increasingly, patients with advanced lung disease are being offered operative procedures. The assessment of the perioperative risk of these patients must include not only the assessment of their lung disease, but the assessment of the patient's cardiovascular disease, their age, and their other medical problems. Knowledge of the stress of particular surgical procedures is also of importance in risk assessment, and is addressed in this article.
Clinics in Chest Medicine 10/1997; 18(3):483-94. · 3.28 Impact Factor
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ABSTRACT: Recent in vivo and in vitro experimental evidence indicates that transforming growth factor-alpha (TGF-alpha) is an important growth factor in the process of recovery and remodeling that occurs after acute lung injury. However, there are very little clinical data on TGF-alpha in patients with acute lung injury. Therefore, the purpose of this study was to determine if TGF-alpha is present in biologically significant concentrations in the pulmonary edema fluid from patients with acute lung injury, and to determine if the presence of TGF-alpha is specific for acute lung injury by including control patients with hydrostatic edema. Using an enzyme-linked immunosorbent assay, plasma and pulmonary edema fluid TGF-alpha levels were measured in 43 patients (34 with increased permeability edema, nine with hydrostatic edema). TGF-alpha was detected in 24 of 34 patients (71%) with increased permeability pulmonary edema (range, 0.035 to 2.57 ng/mL) compared with only two of nine patients with hydrostatic edema (p < 0.05). TGF-alpha was not detected in any plasma samples. These concentrations of TGF-alpha in pulmonary edema fluid have potent in vivo and in vitro effects on alveolar epithelial sodium transport and alveolar epithelial cell motility. In conclusion, biologically relevant concentrations of soluble TGF-alpha are present in the pulmonary edema fluid on day 1 of patients with acute lung injury, a remarkable finding with important implications for the repair and resolution of acute lung injury, particularly since TGF-alpha was detected so early in the course of acute lung injury.
Chest 03/1997; 111(3):652-6. · 5.25 Impact Factor
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ABSTRACT: Adhesive interactions between cells and extracellular matrix proteins are important in cell attachment, migration, and proliferation. The present work defines the role of fibronectin (soluble and insoluble) compared with type I and type IV collagen on in vitro alveolar epithelial wound healing. Repeated video microscopy experiments demonstrated that the half-time of wound closure was decreased in the presence of soluble fibronectin (6.6 +/- 2.1 vs. 17.4 +/- 0.8 h in serum-free medium, P < 0.05). Video microscopy, electron microscopy, and vinculin distribution demonstrated the contribution of two main events during the repair process: the migration of epithelial cell sheets and the spreading of the cells. During the wound healing, the internuclear distance between two adjacent cells at the migrating edge of the wound was significantly increased 10 h after wounding in the presence of soluble fibronectin (67 +/- 3.0 vs. 45 +/- 1.5 microns in serum-free medium, P < 0.05), indicating that cell spreading is involved as part of the mechanism for wound closure. Compared with type I and type IV collagen, insoluble fibronectin was the most potent stimulus for alveolar type II cell motility and wound healing in the absence of other serum factors. These results demonstrate that alveolar epithelial wound healing can be modulated in vitro by the composition of the extracellular matrix, an effect that may be mediated by changes in cell shape.
The American journal of physiology 11/1996; 271(5 Pt 1):L844-53.
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ABSTRACT: Alveolar epithelial type II cells are essential for regenerating an intact alveolar barrier after destruction of type I cells in vivo. The first objective of these experimental studies was to develop an in vitro model to quantify alveolar epithelial cell wound repair. The second objective was to investigate mechanisms of alveolar epithelial cell wound healing by studying the effects of serum and transforming growth factor-alpha (TGF-alpha) on wound closure. Primary cultures of rat alveolar type II cells were prepared by standard methods and grown to form confluent monolayers in 48 h. Then a wound was made by denuding an area (mean initial area of 2.1 +/- 0.6 mm2) of the monolayer. Re-epithelialization of the denuded area over time in the presence or absence of serum was measured using quantitative measurements from time-lapse video microscopy. The half time of wound healing was significantly enhanced in the presence of serum compared with serum-free conditions (2.4 +/- 0.2 vs. 17.4 +/- 0.8 h, P < 0.001). We then tested the hypothesis that TGF-alpha is an important growth factor for stimulating wound repair of alveolar epithelial cells. Exogenous addition of TGF-alpha in serum-free medium resulted in a significantly more rapid wound closure, and, furthermore, the addition of a monoclonal antibody to TGF-alpha in the presence of serum significantly decreased fourfold the rate of wound closure. Measurement of internuclear cell distance confirmed that both cell motility and cell spreading were responsible for closure of the wound. These data demonstrate that 1) the mechanisms of alveolar cell repair can be studied in vitro and that 2) TGF-alpha is a potent growth factor that enhances in vitro alveolar epithelial cell wound closure.
The American journal of physiology 01/1995; 267(6 Pt 1):L728-38.
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ABSTRACT: The effect of hyperosmolar fluid aspiration (seawater) on lung fluid balance has not been well studied. Therefore, the effect of this clinically relevant form of acute lung injury on the alveolar epithelial and lung endothelial barriers was examined in ventilated, anesthetized rabbits. Seawater (4 ml/kg body weight, 881 +/- 29 mOsm/kg) with 3 microCi of 125I-albumin was instilled into the lower trachea of ventilated, anesthetized rabbits. Osmotic equilibration with plasma was completed within the first 5 min after seawater instillation. In parallel with the osmotic equilibration of the seawater in the air spaces, there was a 3-fold dilution of the alveolar protein tracer 125I-albumin, indicating an initial large (300%) increase in alveolar fluid volume. There was a marked decline in arterial oxygenation at the same time that the alveolar fluid volume markedly increased. The initial dilution of the alveolar protein tracer was followed by a progressive increase in the alveolar protein tracer concentration that continued until 6 h after seawater instillation. As the alveolar protein tracer concentrated, arterial oxygenation improved, indicting net alveolar liquid clearance. There was only a mild increase in the epithelial and endothelial permeability to protein within the first 2 h after seawater instillation. Thus, a large osmotically induced increase in alveolar fluid volume with severe pulmonary edema did not cause sustained injury to the endothelial or epithelial barriers of the lung. In fact, normal alveolar liquid clearance occurred, indicating the resistance of the epithelial barrier to hyperosmolar injury as well as its capacity to rapidly reabsorb excess alveolar fluid.
American Journal of Respiratory and Critical Care Medicine 01/1995; 150(6 Pt 1):1555-63. · 11.08 Impact Factor
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ABSTRACT: The movement of water between the air space and capillary compartments is important for the maintenance of air space hydration during respiration and for reabsorption of excess alveolar fluid. We have obtained immunocytochemical and functional evidence that plasma-membrane water channels are responsible for water transport in the intact lung. Northern and quantitative immunoblot analysis showed high expression of CHIP28 (channel-forming integral membrane protein of 28 kDa) water channels in rat lung; immunocytochemistry showed CHIP28 localization to epithelial cell plasma membranes. Stopped-flow light scattering measurements of osmotic water permeability (Pf) in freshly isolated rat alveolar type II epithelial cells indicated a high Pf of 0.015 +/- 0.002 cm/s (10 degrees C) that was weakly temperature-dependent (activation energy, 4 kcal/mol) and reversibly inhibited by 78 +/- 4% by 0.5 mM HgCl2. An in situ-perfused sheep lung model was used to determine the route for water movement in intact lung. Blood-to-air-space water transport was measured by sampling air space fluid after instillation into distal air spaces of hyperosmolar saline (900 mOsm) containing radioiodinated albumin and [14C]mannitol. In seven sets of experiments, air space osmolality and radioiodinated albumin equilibrated with a t1/2 of 0.85 +/- 0.1 min. In the contralateral lung perfused with 0.5 mM HgCl2, t1/2 increased to 2.7 +/- 0.4 min; the inhibitory effect of HgCl2 was fully reversed by 5 mM 2-mercaptoethanol. These results provide direct evidence for transcellular movement of water across the alveolar epithelium in intact lung through mercury-sensitive water channels.
Proceedings of the National Academy of Sciences 06/1994; 91(11):4970-4. · 9.68 Impact Factor
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ABSTRACT: The central importance of the alveolar epithelial barrier in the pathogenesis and recovery from acute lung injury has only recently been appreciated. Both in vivo and in vitro studies have provided a new understanding of the mechanisms that regulate transport of sodium, water, and protein across the alveolar epithelial barrier. This new information regarding the normal function of the alveolar epithelial barrier in regulating lung fluid and protein balance has made it possible to study the function of the alveolar barrier both experimentally and clinically in the setting of acute lung injury. The alveolar epithelial barrier is much more resistant to injury than the nearby lung endothelium. The mechanisms that cause injury to the alveolar barrier are just beginning to be explored in different experimental models of acute lung injury. Some progress has been made in understanding how alveolar barrier injury occurs, especially in bacterial pneumonia. Finally, while it is recognized that alveolar epithelial type II cells play an important role in both ion transport and surfactant production, it is now possible to study in vitro the contribution of alveolar epithelial type II cells in repair of the denuded alveolar barrier.
New horizons (Baltimore, Md.) 12/1993; 1(4):613-22.
