Comparison of Chlamydia trachomatis serovar L2 growth in polarized genital epithelial cells grown in three-dimensional culture with non-polarized cells

Dessus-Babus, Sophie; Moore, Cheryl G.; Whittimore, Judy D.; Wyrick, Priscilla B.

doi:10.1016/j.micinf.2008.02.002

Cited by 9 publications

(7 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although fewer infectious EBs entered polA2EN cells compared to HeLa 229 cells, chlamydiae underwent robust replication that yielded infectious EBs equivalent to numbers of infectious EBs observed in infected HeLa cells (HeLa cell data not shown). These results suggest that the polarized orientation of the A2EN cells creates an intracellular environment that allows for rapid productive replication of chlamydial particles, similar to that reported in polarized endometrial cells [9]. In support of the TEER data, using light microscopy, we observed minimal host cell lysis even after 72 hours in C. trachomatis infected polA2EN cells similar to observations made on C. trachomatis infected trophoblast cells [24].…”

Section: Resultssupporting

confidence: 89%

“…Recent studies, however, have highlighted the importance of the cell type in which chlamydiae are grown, as cell lines derived from different anatomical sites yield different growth rates and infectious yields [9, 10]. Neither HeLa cells nor murine fibroblast cells accurately represent the target cells in vivo , the polarized columnar epithelial cells, with respect to morphology, expression of innate immune mediators or responsiveness to TLR agonists [11, 12].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Chlamydia trachomatis infection results in a modest pro-inflammatory cytokine response and a decrease in T cell chemokine secretion in human polarized endocervical epithelial cells

et al. 2013

View full text Add to dashboard Cite

The endocervical epithelium is a major reservoir for Chlamydia trachomatis in women, and genital infections are extended in their duration. Epithelial cells act as mucosal sentinels by secreting cytokines and chemokines in response to pathogen challenge and infection. We therefore determined the signature cytokine and chemokine response of primary-like endocervix-derived epithelial cells in response to a common genital serovar (D) of C. trachomatis. For these studies, we used a recently-established polarized, immortalized, endocervical epithelial cell model (polA2EN) that maintains, in vitro, the architectural and functional characteristics of endocervical epithelial cells in vivo including the production of pro-inflammatory cytokines. PolA2EN cells were susceptible to C. trachomatis infection, and chlamydiae in these cells underwent a normal developmental cycle as determined by a one-step growth curve. IL1α protein levels were increased in both apical and basolateral secretions of C. trachomatis infected polA2EN cells, but this response did not occur until 72 hours after infection. Furthermore, protein levels of the pro-inflammatory cytokines and chemokines IL6, TNFα and CXCL8 were not significantly different between C. trachomatis infected polA2EN cells and mock infected cells at any time during the chlamydial developmental cycle up to 120 hours post-infection. Intriguingly, C. trachomatis infection resulted in a significant decrease in the constitutive secretion of T cell chemokines IP10 and RANTES, and this required a productive C. trachomatis infection. Examination of anti-inflammatory cytokines revealed a high constitutive apical secretion of IL1ra from polA2EN cells that was not significantly modulated by C. trachomatis infection. IL-11 was induced by C. trachomatis, although only from the basolateral membrane. These results suggest that C. trachomatis can use evasion strategies to circumvent a robust pro-inflammatory cytokine and chemokine response. These evasion strategies, together with the inherent immune repertoire of endocervical epithelial cells, may aid chlamydiae in establishing, and possibly sustaining, an intracellular niche in microenvironments of the endocervix in vivo.

show abstract

Section: Resultssupporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Chlamydia trachomatis infection results in a modest pro-inflammatory cytokine response and a decrease in T cell chemokine secretion in human polarized endocervical epithelial cells

et al. 2013

View full text Add to dashboard Cite

show abstract

“…In a typical serovar L2 developmental cycle (organisms grown in HeLa or HEp2 cells), rapid division of RBs occurs between 8 and 16 h post-infection, with RB to EB differentiation occurring 24 to 36 h post-infection. Maximal RB to EB transition occurs between 42 and 48 h post-infection, with subsequent monolayer destruction due to maximal EB release occurring at or after 50+ h of infection (Ward, 1988 ; Dessus-Babus et al, 2008 ). Consistent with the progression of the normal chlamydial developmental cycle, there are low numbers of infectious progeny produced in both siRNA treatment groups (NT: 3.54 × 10 4 ± 3.97 × 10 3 ; Stx10: 2.41 × 10 4 ± 2.24 × 10 3 ) after 24 h of infection (mid-developmental cycle), indicating that chlamydial development is not altered due to depletion of syntaxin 10 at this time point post-infection.…”

Section: Resultsmentioning

confidence: 99%

The trans-Golgi SNARE syntaxin 10 is required for optimal development of Chlamydia trachomatis

Lucas

Ouellette

Kabeiseman

et al. 2015

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

Chlamydia trachomatis, an obligate intracellular pathogen, grows inside of a vacuole, termed the inclusion. Within the inclusion, the organisms differentiate from the infectious elementary body (EB) into the reticulate body (RB). The RB communicates with the host cell through the inclusion membrane to obtain the nutrients necessary to divide, thus expanding the chlamydial population. At late time points within the developmental cycle, the RBs respond to unknown molecular signals to redifferentiate into infectious EBs to perpetuate the infection cycle. One strategy for Chlamydia to obtain necessary nutrients and metabolites from the host is to intercept host vesicular trafficking pathways. In this study we demonstrate that a trans-Golgi soluble N-ethylmaleimide–sensitive factor attachment protein (SNARE), syntaxin 10, and/or syntaxin 10-associated Golgi elements colocalize with the chlamydial inclusion. We hypothesized that Chlamydia utilizes the molecular machinery of syntaxin 10 at the inclusion membrane to intercept specific vesicular trafficking pathways in order to create and maintain an optimal intra-inclusion environment. To test this hypothesis, we used siRNA knockdown of syntaxin 10 to examine the impact of the loss of syntaxin 10 on chlamydial growth and development. Our results demonstrate that loss of syntaxin 10 leads to defects in normal chlamydial maturation including: variable inclusion size with fewer chlamydial organisms per inclusion, fewer infectious progeny, and delayed or halted RB-EB differentiation. These defects in chlamydial development correlate with an overabundance of NBD-lipid retained by inclusions cultured in syntaxin 10 knockdown cells. Overall, loss of syntaxin 10 at the inclusion membrane negatively affects Chlamydia. Understanding host machinery involved in maintaining an optimal inclusion environment to support chlamydial growth and development is critical toward understanding the molecular signals involved in successful progression through the chlamydial developmental cycle.

show abstract

“…We found a similar, clustered appearance of C. pneumoniae inclusions in both the polarized and the flat Calu-3 cultures but not in either of the A549 cultures (results not shown). This phenomenon should be further studied in order to determine whether there are certain receptor molecules enriched on various areas of an apical membrane where some chlamydia serovars or species are able to attach, as suggested by Dessus-Babus et al (1).…”

Section: Discussionmentioning

confidence: 99%

Chlamydia pneumoniae Infection in Polarized Epithelial Cell Lines

Törmäkangas

Markkula

Lounatmaa³

et al. 2010

Infect Immun

View full text Add to dashboard Cite

We set up a polarized cell culture model to study the pathogenicity of a common respiratory tract pathogen, Chlamydia pneumoniae. Immunofluorescence staining of ZO-1 (a tight junction protein) and Na ؉ K ؉ ATPase (a protein pump localized at the basolateral membrane in the polarized epithelial cells), as well as TER measurements, suggested that the filter-grown Calu-3 cells, but not the A549 cells, were polarized when grown on collagen-coated membranes. Both the flat and the filter-grown cultures were infected with C. pneumoniae. Infection in the polarized Calu-3 cultures produced more C. pneumoniae genome equivalents than infection in the flat cultures. However, this progeny was not as infective as that in the flat cultures. The maximum amount of C. pneumoniae was detected at 6 days postinfection in the filter-grown A549 cells, indicating a slower developmental cycle than that observed in the flat A549 cultures. The effect of cycloheximide on the growth of C. pneumoniae in the polarized cells was negligible. Furthermore, the infection in the polarized Calu-3 cells was resistant to doxycycline, and several cytokines were released mainly on the apical side of the polarized cells in response to C. pneumoniae infection. These findings indicate that the growth of chlamydiae was altered in the filter-grown epithelial culture system. The diminished production of infective progeny of C. pneumoniae, together with the resistance to doxycycline and polarized secretion of cytokines from the infected Calu-3 cells, suggests that this model is useful for examining epithelial cell responses to C. pneumoniae infection, and it might better resemble in vivo infection in respiratory epithelial cells.

show abstract

Comparison of Chlamydia trachomatis serovar L2 growth in polarized genital epithelial cells grown in three-dimensional culture with non-polarized cells

Cited by 9 publications

References 25 publications

Chlamydia trachomatis infection results in a modest pro-inflammatory cytokine response and a decrease in T cell chemokine secretion in human polarized endocervical epithelial cells

Chlamydia trachomatis infection results in a modest pro-inflammatory cytokine response and a decrease in T cell chemokine secretion in human polarized endocervical epithelial cells

The trans-Golgi SNARE syntaxin 10 is required for optimal development of Chlamydia trachomatis

Chlamydia pneumoniae Infection in Polarized Epithelial Cell Lines

Contact Info

Product

Resources

About