Use of green fluorescent protein in visualisation of pneumococcal invasion of broncho-epithelial cells in vivo

Kadioglu, Aras

doi:10.1016/s0378-1097(00)00497-3

Cited by 11 publications

(13 citation statements)

References 26 publications

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“…This study presents data on one particular system, TCS09, that is important in virulence in two different strains of the pneumococcus, D39 and 0100993. Control studies using D39 transformed with plasmid pVA838, which does not integrate into the chromosome, have shown that the virulence of this strain is not compromised by the transformation procedure used to create the mutants, confirming observations made by Kadioglu et al (15). Administration of S. pneumoniae via the intraperitoneal route generally results in high levels of bacteremia within 24 h of injection together with the display of moderate to severe symptoms (hunched stance, piloerection, lethargy) which progress until the mice become moribund.…”

Section: Discussionsupporting

confidence: 86%

Contribution of a Response Regulator to the Virulence of Streptococcus pneumoniae Is Strain Dependent

Blue

Mitchell

2003

Infect Immun

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Bacterial two-component signal transduction systems (TCS) enable bacteria to respond to environmental changes and regulate a range of genes accordingly. They have a crucial role in regulating many cellular responses and have excellent potential as antibacterial-drug targets. We have constructed mutations in a TCS response regulator gene for two different strains of the human pathogen Streptococcus pneumoniae. These mutants have been analyzed in our murine model of infection. Data suggest that in a D39 background the response regulator gene is essential for virulence; an isogenic mutant is avirulent via intraperitoneal, intranasal, and intravenous routes of infection. This mutant, which does not show impaired growth in vitro, is unable to grow in the lung tissue or in blood. Mutation of the response regulator in a 0100993 background results in a strain that is fully virulent intraperitoneally and intravenously but shows decreased levels of bacteremia and increased murine survival following intranasal infection. The ability to grow in the lung tissue is not impaired in this mutant, suggesting that it has an impaired ability to disseminate from the lungs to the systemic circulation. Our data highlight the importance of assessing the contribution of putative virulence factors to the infection process at different sites of infection and provide evidence that virulence determinants can behave very differently based on the genetic background of the bacterial strain. These important findings may be relevant to other bacterial pathogens.

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Section: Discussionsupporting

confidence: 86%

Contribution of a Response Regulator to the Virulence of Streptococcus pneumoniae Is Strain Dependent

Blue

Mitchell

2003

Infect Immun

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“…However, confirming that the bacteria invade mesothelial cells during infection in mice would require electron or confocal microscopy of recovered mesothelium and would be technically very challenging due to the low number of events. Several mechanisms of cell invasion dependent on different bacterial protein/host ligand interactions have been described for S. pneumoniae and can result in translocation across epithelial and endothelial layers (21,(26)(27)(28)(29). Further research is required to clarify whether similar mechanisms are relevant for translocation across mesothelial cells, characterization of which could offer potential targets for therapies that prevent empyema.…”

Section: Discussionmentioning

confidence: 99%

Characterization of a New Mouse Model of Empyema and the Mechanisms of Pleural Invasion by Streptococcus pneumoniae

Wilkosz

Edwards

Bielsa

et al. 2012

Am J Respir Cell Mol Biol

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Although empyema affects more than 65,000 people each year in the United States and in the United Kingdom, there are limited data on the pathogenesis of pleural infection. We investigated the pathogenesis of empyema using animal and cell culture models of Streptococcus pneumoniae infection. The pathological processes during the development of empyema associated with murine pneumonia due to S. pneumoniae (strain D39) were investigated. Lungs were examined using histology, and pleural fluid and blood bacterial colony-forming units, cytokine levels, and cellular infiltrate were determined over time. Bacterial migration across mesothelial monolayers was investigated using cell culture techniques, flow cytometry, and confocal microscopy. After intranasal inoculation with 10 7 S. pneumoniae D39 strain, mice developed pneumonia associated with rapid bacterial invasion of the pleural space; raised intrapleural IL-8, VEGF, MCP-1, and TNF-a levels; and caused significant intrapleural neutrophilia followed by the development of fibrinous pleural adhesions. Bacterial clearance from the pleural space was poor, and in vitro assays demonstrated that S. pneumoniae crossed mesothelial layers by translocation through cells rather than by a paracellular route. This study describes key events during the development of S. pneumoniae empyema using a novel murine model of pneumonia-associated empyema that closely mimics human disease. The model allows for future assessment of molecular mechanisms involved in the development of empyema and evaluation of potential new therapies. The data suggest that transmigration of bacteria through mesothelial cells could be important in empyema development. Furthermore, upon entry the pleural cavity offers a protected compartment for the bacteria.Keywords: Streptoccocus pneumoniae; empyema; animal models; translocation; mesothelial cells Pleural infection is a global problem that affects over 65,000 patients each year in the United States and in the United Kingdom and is associated with significant morbidity and mortality (1-5). Empyema is defined by the presence of bacteria or pus in the pleural cavity and usually develops as a complication of pneumonia. Important questions regarding the disease pathogenesis remain unanswered, which may in part account for the lack of recent therapeutic advances. A key limiting factor in this area of research is that there is no suitable preclinical animal model of empyema. Studying the evolution of pleuropulmonary infection ideally involves repeated sampling of the lung and pleural cavity, which is not feasible in patients with pneumonia, and the animal models used to date have significant limitations (6). Published in vivo models rely on intrapleural delivery of bacteria to create localized pleural infection without concurrent pneumonia and many use species-specific microorganisms, such as Pasteurella multocida (7-9), with limited relevance to humans. These models require concomitant administration of systemic antibiotics to control infection and fatalities (8), ...

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“…Of particular relevance is the type 2 D39 reference strain (142), which several research groups have employed to study the VOL. 21,2008 MODELS OF PNEUMOCOCCAL DISEASE 667 pathogenesis of pneumococcal disease and test vaccine antigens over the years (25,40,50,92,103,104,109,123,124,131,(180)(181)(182)186). Serotype 3 strains, in particular A66 (9) and WU2 (43), are also widely used for inducing sepsis in the mouse and the rat (2,40,43,109,129,163) and to study invasive pneumococcal disease in the rabbit (246,247).…”

Section: Considerations Of Animal and Pneumococcal Strainsmentioning

confidence: 99%

“…infected animals were inoculated into the nostrils of anesthetized mice, which rapidly developed bronchopneumonia with bacteremia (50). This model has been employed largely in studies on host immune response to pneumococcal lung infection (23,70,122,123,130), mouse susceptibility and resistance to the disease (92), invasion by S. pneumoniae of bronchoepithelial cells in vivo (124), the role of virulence determinants in pneumonia (30,58,120,125,158), and the efficacy of antibiotics and anti-inflammatory drugs (250,252). Furthermore, many investigations on protection from i.n.…”

Section: Mouse Modelsmentioning

confidence: 99%

Animal Models ofStreptococcus pneumoniaeDisease

2008

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SUMMARY Streptococcus pneumoniae is a colonizer of human nasopharynx, but it is also an important pathogen responsible for high morbidity, high mortality, numerous disabilities, and high health costs throughout the world. Major diseases caused by S. pneumoniae are otitis media, pneumonia, sepsis, and meningitis. Despite the availability of antibiotics and vaccines, pneumococcal infections still have high mortality rates, especially in risk groups. For this reason, there is an exceptionally extensive research effort worldwide to better understand the diseases caused by the pneumococcus, with the aim of developing improved therapeutics and vaccines. Animal experimentation is an essential tool to study the pathogenesis of infectious diseases and test novel drugs and vaccines. This article reviews both historical and innovative laboratory pneumococcal animal models that have vastly added to knowledge of (i) mechanisms of infection, pathogenesis, and immunity; (ii) efficacies of antimicrobials; and (iii) screening of vaccine candidates. A comprehensive description of the techniques applied to induce disease is provided, the advantages and limitations of mouse, rat, and rabbit models used to mimic pneumonia, sepsis, and meningitis are discussed, and a section on otitis media models is also included. The choice of appropriate animal models for in vivo studies is a key element for improved understanding of pneumococcal disease.

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Use of green fluorescent protein in visualisation of pneumococcal invasion of broncho-epithelial cells in vivo

Cited by 11 publications

References 26 publications

Contribution of a Response Regulator to the Virulence of Streptococcus pneumoniae Is Strain Dependent

Contribution of a Response Regulator to the Virulence of Streptococcus pneumoniae Is Strain Dependent

Characterization of a New Mouse Model of Empyema and the Mechanisms of Pleural Invasion by Streptococcus pneumoniae

Animal Models ofStreptococcus pneumoniaeDisease

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