Gonococcal and meningococcal pathogenesis as defined by human cell, cell culture, and organ culture assays

Stephens, David S.

doi:10.1128/cmr.2.suppl.s104

Cited by 49 publications

(36 citation statements)

References 60 publications

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“…The data demonstrated that a sustained, albeit low-level, C. trachomatis infection occurs in the niodel. In contrast to gonococcal infection of HFTOC, where lipopolysaccharides and peptidoglycan monomers released from rapidly dividing gonococci extensively damage both ciliated and nonciliated epithelial cells (Stephens, 1989), infection of HFTOC with C. trachomat u had no overall effect on mucosal architecture or ciliary activity. These data support the results of Hutchinson et al (1979), who found that ciliary activity of fallopian tube mucosa was not diminished by C. trachomatis infection.…”

Section: Discussionmentioning

confidence: 87%

Chlamydia trachomatis infection of human fallopian tube organ cultures

Cooper

Rapp²,

Jeffery-Wiseman³

et al. 1990

Journal of General Microbiology

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The pathogenic events that precede Chlamydia trachomatis salpingitis in the human fallopian tube have not been fully described. We used a model of human fallopian tubes in organ culture (HFTOC) infected with strain E/UW-5/CX of C. trachomatis to study these events. The model supported sustained C. trachomatis infection as demonstrated by recovery of viable C. trachomatis from medium and tissue over 5-7 d. However, the level of infectivity was low. Maximal infection occurred at 72 h after initial inoculation. In contrast to gonococcal infection of the HFTOC, C. trachomatis did not damage overall ciliary function of HFTOC. However, a local direct cytotoxic effect characterized by loss of microvilli and disruption of cell junctions was noted when multiple chlamydial elementary bodies attached to mucosal cells. Beginning at 24 h, and continuing throughout the course of C. trachomatis infection of HFTOC, ruptured epithelial cells releasing elementary bodies were noted. Chlamydia1 inclusions were seen in the mucosa by 72 h in -6% of both ciliated and nonciliated epithelial cells. Mucosal inclusions contained all forms of the C. trachomatis developmental cycle. These data suggest that factors present in the human fallopian tube may limit susceptibility to chlamydial infection but support the use of the HFTOC model in the study of the pathogenesis of C. trachomatis salpingitis.

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Section: Discussionmentioning

confidence: 87%

Chlamydia trachomatis infection of human fallopian tube organ cultures

Cooper

Rapp²,

Jeffery-Wiseman³

et al. 1990

Journal of General Microbiology

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“…A number of microscopy studies have explored the ultrastructure of the N. gonorrhoeae-infected epithelial cell 3D ultrastructure of N. gonorrhoeae infection cortex and the bacterial microcolony (Edwards et al, 2000;Evans, 1977;McGee et al, 1983;Steichen et al, 2008;Stephens, 1989;Swanson, 1983;Ward & Watt, 1972). GC are often seen to attach as microcolonies and to induce dramatic alterations to host cell microvilli, particularly at the microcolony periphery.…”

Section: Discussionmentioning

confidence: 99%

“…Microvilli beneath GC microcolonies are numerous and appear elongated (Evans, 1977;Griffiss et al, 1999;McGee et al, 1981;Mosleh et al, 1997;Stephens, 1989;Tjia et al, 1988;Ward et al, 1974). We examined GC-infected cells by SEM to determine the role of Tfp in microvillus architecture.…”

Section: Sem Of the Gc-infected Cell Cortexmentioning

confidence: 99%

“…The relationship between the cortical plaque and the microcolony is dynamic. As the microcolony crawls over the cell surface, the cortical plaque migrates with it (Higashi et al, 2007).The microcolony-host cell interface is therefore an important focus of study.A number of microscopy studies have provided important information on the ultrastructure of the infected host cell surface (Edwards et al, 2000;Evans, 1977;Griffiss et al, 1999;McGee et al, 1981;Mosleh et al, 1997;Stephens, 1989;Tjia et al, 1988;Ward et al, 1974). Microvilli are numerous at sites of infection, and they appear to be elongated and deformed.…”

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confidence: 99%

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Influence of type IV pilus retraction on the architecture of the Neisseria gonorrhoeae-infected cell cortex

et al. 2009

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Early in infection, Neisseria gonorrhoeae can be observed to attach to the epithelial cell surface as microcolonies and induce dramatic changes to the host cell cortex. We tested the hypothesis that type IV pili (Tfp) retraction plays a role in the ultrastructure of both the host cell cortex and the bacterial microcolony. Using serial ultrathin sectioning, transmission electron microscopy and 3D reconstruction of serial 2D images, we have obtained what we believe to be the first 3D reconstructions of the N. gonorrhoeae-host cell interface, and determined the architecture of infected cell microvilli as well as the attached microcolony. Tfp connect both wild-type (wt) and Tfp retraction-deficient bacteria with each other, and with the host cell membrane. Tfp fibres and microvilli form a lattice in the wt microcolony and at its periphery. Wt microcolonies induce microvilli formation and increases of surface area, leading to an approximately ninefold increase in the surface area of the host cell membrane at the site of attachment. In contrast, Tfp retractiondeficient microcolonies do not affect these parameters. Wt microcolonies had a symmetrical, dome-shaped structure with a circular 'footprint', while Tfp retraction-deficient microcolonies were notably less symmetrical. These findings support a major role for Tfp retraction in microvilli and microcolony architecture. They are consistent with the biophysical attributes of Tfp and the effects of Tfp retraction on epithelial cell signalling. INTRODUCTIONNeisseria gonorrhoeae (the gonococcus, or GC) remains a significant public health concern. The Gram-negative diplococcus causes a million cases of gonorrhoea annually in Western Europe and North America, and over 62 million cases worldwide (World Health Organization). Humans are the only known reservoir for GC. The exquisite tropism of GC for man and the ability of GC to establish a carrier state (Turner et al., 2002) reflect a highly evolved relationship between pathogen and host.Gonococcal attachment is promoted by type IV pili (Tfp), peritrichous fibres that are 6 nm in width and up to several micrometres in length. The Tfp filament is composed of helical polymers of pilin subunits (Craig et al., 2006;Parge et al., 1995) and minor proteins (Carbonnelle et al., 2005;Winther-Larsen et al., 2005). Tfp participate in important biological processes such as DNA uptake/genetic exchange, twitching motility, attachment, and host cell signalling (Mattick, 2002;. These activities require, or are enhanced by, the retraction of Tfp fibres (Wolfgang et al., 2000).Gonococcal Tfp retraction is an activity that requires PilT, the Tfp retraction motor subunit (Wolfgang et al., 1998). Retracting Tfp generate strong 'pull' forces, ranging from 50 picoNewtons (pN) to 1 nanoNewton (nN) (Biais et al., 2008;Opitz et al., 2009). Repeated cycles of Tfp extension, substrate attachment and Tfp retraction allow GC to crawl over surfaces and aggregate into microcolonies. Tfp retraction from bacteria in a microcolony allows the community to craw...

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“…These two events may be mediated by single or multiple microbial factors (1). Attempts to sort out the role of putative attachment and invasion factors of Neisreriagonorrhoerze (GC) have utilized a variety of cell types and models, many of which use resistance to killing by extracellular agents, such as gentamicin, to distinguish extracellular from intracellular bacteria (2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13). Results of these investigations have not been uniform with different cell types (6,12,13), so extrapolation of these data to infections of natural target tissues is difficult.…”

Section: Introductionmentioning

confidence: 99%

Digital confocal microscopy allows measurement and three-dimensional multiple spectral reconstruction of Neisseria gonorrhoeae/epithelial cell interactions in the human fallopian tube organ culture model.

Gorby

1994

J Histochem Cytochem.

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A strategy for measuring Neiseria gonorrhoeae attachment and invasion in the human Fallopian tube organ culture ( m c ) model via computerized image analysis (CIA) combined with "digital" confocal microscopy (DCM) was tested. DCM on serial image stacks of fluorescent latex beads reduced out-of-focus light propagation in theZ-axis @<0.005) and improved the shape factor of lateral three-dimensional reconstructions of the beads @<0.001). Sections of tissue infected for 44 hr with piliated, Opt+ gonococci were stained with fluorescein-labeled monoclonal a n t i -g o n d antibodies, rhodamine-labeled phalloidin, and Hoechst 33342. Serial images collected at identical focal planes for each fluorochrome were subjected to DCM. Epithelial cytoplasmic regions of interest defied by rhodamine-stained actin

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Gonococcal and meningococcal pathogenesis as defined by human cell, cell culture, and organ culture assays

Cited by 49 publications

References 60 publications

Chlamydia trachomatis infection of human fallopian tube organ cultures

Chlamydia trachomatis infection of human fallopian tube organ cultures

Influence of type IV pilus retraction on the architecture of the Neisseria gonorrhoeae-infected cell cortex

Digital confocal microscopy allows measurement and three-dimensional multiple spectral reconstruction of Neisseria gonorrhoeae/epithelial cell interactions in the human fallopian tube organ culture model.

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