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Identification of meningococcal genes necessary for colonization of human upper airway tissue.

ABSTRACT: Neisseria meningitidis is an exclusively human pathogen that has evolved primarily to colonize the nasopharynx rather than to cause systemic disease. Colonization is the most frequent outcome following meningococcal infection and a prerequisite for invasive disease. The mechanism of colonization involves attachment of the organism to epithelial cells via bacterial type IV pili (Tfp), but subsequent events during colonization remain largely unknown. We analyzed 576 N. meningitidis mutants for their capacity to colonize human nasopharyngeal tissue in an organ culture model to identify bacterial genes required for colonization. Eight colonization-defective mutants were isolated. Two mutants were unable to express Tfp and were defective for adhesion to epithelial cells, which is likely to be the basis of their attenuation in nasopharyngeal tissue. Three other mutants are predicted to have lost previously uncharacterized surface molecules, while the remaining mutants have transposon insertions in genes of unknown function. We have identified novel meningococcal colonization factors, and this should provide insights into the survival of this important pathogen in its natural habitat.

SUBMITTER: Exley RM

PROVIDER: S-EPMC2612245 | biostudies-literature | 2009 Jan

REPOSITORIES: biostudies-literature

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Identification of meningococcal genes necessary for colonization of human upper airway tissue.

Exley Rachel M RM Sim Richard R Goodwin Linda L Winterbotham Megan M Schneider Muriel C MC Read Robert C RC Tang Christoph M CM

Infection and immunity 20081020 1

Neisseria meningitidis is an exclusively human pathogen that has evolved primarily to colonize the nasopharynx rather than to cause systemic disease. Colonization is the most frequent outcome following meningococcal infection and a prerequisite for invasive disease. The mechanism of colonization involves attachment of the organism to epithelial cells via bacterial type IV pili (Tfp), but subsequent events during colonization remain largely unknown. We analyzed 576 N. meningitidis mutants for the ...[more]

PMID: 18936183

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Project description:UNLABELLED:Neisseria meningitidis is a leading cause of bacterial meningitis and septicemia, affecting infants and adults worldwide. N. meningitidis is also a common inhabitant of the human nasopharynx and, as such, is highly adapted to its niche. During bacteremia, N. meningitidis gains access to the blood compartment, where it adheres to endothelial cells of blood vessels and causes dramatic vascular damage. Colonization of the nasopharyngeal niche and communication with the different human cell types is a major issue of the N. meningitidis life cycle that is poorly understood. Here, highly saturated random transposon insertion libraries of N. meningitidis were engineered, and the fitness of mutations during routine growth and that of colonization of endothelial and epithelial cells in a flow device were assessed in a transposon insertion site sequencing (Tn-seq) analysis. This allowed the identification of genes essential for bacterial growth and genes specifically required for host cell colonization. In addition, after having identified the small noncoding RNAs (sRNAs) located in intergenic regions, the phenotypes associated with mutations in those sRNAs were defined. A total of 383 genes and 8 intergenic regions containing sRNA candidates were identified to be essential for growth, while 288 genes and 33 intergenic regions containing sRNA candidates were found to be specifically required for host cell colonization. IMPORTANCE:Meningococcal meningitis is a common cause of meningitis in infants and adults. Neisseria meningitidis (meningococcus) is also a commensal bacterium of the nasopharynx and is carried by 3 to 30% of healthy humans. Under some unknown circumstances, N. meningitidis is able to invade the bloodstream and cause either meningitis or a fatal septicemia known as purpura fulminans. The onset of symptoms is sudden, and death can follow within hours. Although many meningococcal virulence factors have been identified, the mechanisms that allow the bacterium to switch from the commensal to pathogen state remain unknown. Therefore, we used a Tn-seq strategy coupled to high-throughput DNA sequencing technologies to find genes for proteins used by N. meningitidis to specifically colonize epithelial cells and primary brain endothelial cells. We identified 383 genes and 8 intergenic regions containing sRNAs essential for growth and 288 genes and 33 intergenic regions containing sRNAs required specifically for host cell colonization.

Project description:Study objectivesReal-time monitoring of upper airway collapse during sleep could be instrumental for studies in biomechanics of obstructive sleep apnea (OSA) and selecting individualized treatment modalities. Although some imaging techniques are used under sedated sleep, none are available during the entire natural sleep process. We hypothesized that electrical impedance tomography (EIT) can be used for noninvasive continuous imaging of the upper airway during natural sleep and quantifying upper airway collapse in terms of its size.MethodsAfter determining surface landmarks to attach the electrodes for monitoring the retroglossal airway, EIT was conducted in 10 healthy participants. As a feasibility test of EIT in detecting upper airway collapse, transient airway closure was induced by the swallowing maneuver. These EIT images were confirmed by simultaneous magnetic resonance imaging (MRI) scans. Subsequently, EIT scans were conducted in 7 healthy participants and 10 patients with OSA under nonsedated sleep to determine whether it could identify upper airway narrowing or collapse. Respiratory events were identified by concurrent polysomnography (PSG).ResultsSwallowing-induced airway closure was identified successfully in all 10 participants on simultaneous EIT and MRI scans. Sizes and positions of the upper airway closures in reconstructed EIT images were well correlated with those in magnetic resonance images. Obstructive hypopnea and apnea were detected successfully by EIT in 10 patients with OSA, and no significant changes in EIT data were observed in 7 healthy participants during concurrent EIT and PSG tests. Additionally, conductivity changes in the airway were greater during obstructive apnea than during hypopnea (64.3% versus 26.3%, respectively; P < .001) compared with those during baseline respiration.ConclusionsEIT could be a useful real-time monitoring device for detecting upper airway narrowing or collapse during natural sleep in patients with OSA. Currently, changes in the upper airway size can be estimated with good accuracy, but shape estimation needs future improvements in the EIT image quality.

Dataset Information

Identification of meningococcal genes necessary for colonization of human upper airway tissue.

Publications

Identification of meningococcal genes necessary for colonization of human upper airway tissue.

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