Characterization of estrogen-responsive epithelial cell lines and their infectivity by genital Chlamydia trachomatis

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

doi:10.1016/j.micinf.2005.05.004

Cited by 19 publications

(21 citation statements)

References 46 publications

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“…Alterations of chlamydial protein secretion may allow C. trachomatis to achieve intracellular survival upon ampicillin exposure, therefore protecting the bacteria from innate and acquired immunity. Our in vitro primary human endocervical epithelial cell model provides a valuable tool to investigate host cell factors that are relevant to the establishment of persistence, as this model most likely retains the appropriate responsiveness to endogenous factors, such as female steroid hormones that can be altered in transformed cervical cell lines (Guseva et al, 2005). While this study has focused on selective chlamydial secretable proteins, a broader proteomic analysis is warranted to elucidate how the overall secretome affects the infection process.…”

Section: Discussionmentioning

confidence: 99%

Altered protein secretion of Chlamydia trachomatis in persistently infected human endocervical epithelial cells

et al. 2011

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Chlamydia trachomatis is the most common bacterial infection of the human reproductive tract globally; however, the mechanisms underlying the adaptation of the organism to its natural target cells, human endocervical epithelial cells, are not clearly understood. To secure its intracellular niche, C. trachomatis must modulate the host cellular machinery by secreting virulence factors into the host cytosol to facilitate bacterial growth and survival. Here we used primary human endocervical epithelial cells and HeLa cells infected with C. trachomatis to examine the secretion of bacterial proteins during productive growth and persistent growth induced by ampicillin. Specifically, we observed a decrease in secretable chlamydial protease-like activity factor (CPAF) in the cytosol of host epithelial cells exposed to ampicillin with no evident reduction of CPAF product by C. trachomatis. In contrast, the expression of CopN and Tarp was downregulated, suggesting that C. trachomatis responds to ampicillin exposure by selectively altering the expression of secretable proteins. In addition, we observed a greater accumulation of outer-membrane vesicles from C. trachomatis in persistently infected cells. Taken together, these results suggest that the regulation of both gene expression and the secretion of chlamydial virulence proteins is involved in the adaptation of the bacteria to a persistent infection state in human genital epithelial cells.

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Section: Discussionmentioning

confidence: 99%

Altered protein secretion of Chlamydia trachomatis in persistently infected human endocervical epithelial cells

et al. 2011

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“…This intriguing difference between strains may possibly be due to the recognition of and binding to different receptor molecules on host cell surfaces that, for serovar E, may be enriched or clustered in some regions or microdomains of polarized epithelial cell apical membranes, such as lipid rafts or caveolae, proposed to be involved in serovar E but not in serovar L2 entry [19,20]. Although their role in chlamydial entry is still controversial [21][22][23], it is an interesting possibility, as many pathogens are known to interact with these membrane microdomains and that receptor molecules, such as membrane-associated estrogen receptors that locate to caveolae, have been implicated in serovar E attachment/entry [11,24]. In contrast, initial interactions of serovar L2 EB with host cell surfaces may occur via recognition of a broader range of host molecules or molecules widely represented throughout cell surfaces, such as heparan sulfate proteoglycans [25], and/or via Tarp-mediated pedestallike formation [26,27], a phenomenon shown to be more prevalent for serovar L2 than with serovar D [26] or serovar E [28].…”

Section: Discussionmentioning

confidence: 99%

“…For instance, C. trachomatis serovar E EB progeny recovered from McCoy cell fibroblasts grown on collagen-coated microcarrier beads in a 3D culture system were higher in numbers and more infectious on a per particle ratio basis, because of an accelerated developmental cycle, compared to the progeny collected from McCoy cells grown in flasks [9,10]; in addition, the harvested chlamydiae always exhibited significantly higher infectious titers in more relevant genital epithelial cells than in McCoy cells. An epithelial cell environment appears important for optimal chlamydial growth but the anatomical origin of the cell lines used seems to be critical as well, as emphasized by recent studies [11,12]. For instance, Miyairi et al [12] showed that cell lines originating from different anatomical sites, e.g.…”

Section: Introductionmentioning

confidence: 99%

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

Dessus-Babus

Moore

Whittimore

et al. 2008

Microbes and Infection

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A common model for studying Chlamydia trachomatis and growing chlamydial stocks uses Lymphogranuloma venereum serovar L2 and non-polarized HeLa cells. However, recent publications indicate that the growth rate and progeny yields can vary considerably for a particular strain depending on the cell line/type used, and seem to be partially related to cell tropism. In the present study, the growth of invasive serovar L2 was compared in endometrial HEC-1B and endocervical HeLa cells polarized on collagen-coated microcarrier beads, as well as in HeLa cells grown in tissue culture flasks. Microscopy analysis revealed no difference in chlamydial attachment/ entry patterns or in inclusion development throughout the developmental cycle between cell lines. Very comparable growth curves in both cell lines were also found using real-time PCR analysis, with increases in chlamydial DNA content of 400-500-fold between 2 and 36 h post-inoculation. Similar progeny yields with comparable infectivity were recovered from HEC-1B and HeLa cell bead cultures, and no difference in chlamydial growth was found in polarized vs. non-polarized HeLa cells. In conclusion, unlike other C. trachomatis strains such as urogenital serovar E, invasive serovar L2 grows equally well in physiologically different endometrial and endocervical environments, regardless of the host cell polarization state.

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“…Two micrograms of total RNA from all bead culture samples was then reverse transcribed using the SuperScript II kit (Invitrogen, Carlsbad, CA) in the presence of 200 ng of random hexamers (Integrated DNA Technologies, Coralville, IA) under the conditions recommended by the manufacturer. Prior to qPCR, the quality of all cDNA was checked by conventional PCR with primers specific for human aldolase as previously described (21). In addition, for each RNA sample, an "RT-minus" control, in which the reverse transcriptase (RT) enzyme was omitted, was also included.…”

Section: Vol 75 2007 C Trachomatis Growth Varies In Polarized Epitmentioning

confidence: 99%

“…However, chlamydial infection in various polarized epithelial cell lines also differs. C. trachomatis serovar E infectivity is greater for the endometrial carcinoma epithelial subclone HEC-1B than for the Ishikawa endometrial epithelial cell line and the two breast cancer epithelial cell lines MCF-7 and HCC-1806 (21).…”

mentioning

confidence: 91%

Differences in Chlamydia trachomatis Serovar E Growth Rate in Polarized Endometrial and Endocervical Epithelial Cells Grown in Three-Dimensional Culture

et al. 2007

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In vitro studies of obligate intracellular chlamydia biology and pathogenesis are highly dependent on the use of experimental models and growth conditions that mimic the mucosal architecture and environment these pathogens encounter during natural infections. In this study, the growth of Chlamydia trachomatis genital serovar E was monitored in mouse fibroblast McCoy cells and compared to more relevant host human epithelial endometrium-derived HEC-1B and cervix-derived HeLa cells, seeded and polarized on collagencoated microcarrier beads, using a three-dimensional culture system. Microscopy analysis of these cell lines prior to infection revealed morphological differences reminiscent of their in vivo architecture. Upon infection, early chlamydial inclusion distribution was uniform in McCoy cells but patchy in both epithelial cell lines. Although no difference in chlamydial attachment to or entry into the two genital epithelial cell lines was noted, active bacterial genome replication and transcription, as well as initial transformation of elementary bodies to reticulate bodies, were detected earlier in HEC-1B than in HeLa cells, suggesting a faster growth, which led to higher progeny counts and titers in HEC-1B cells upon completion of the developmental cycle. Chlamydial development in the less relevant McCoy cells was very similar to that in HeLa cells, although higher progeny counts were obtained. In conclusion, this three-dimensional bead culture system represents an improved model for harvesting large quantities of infectious chlamydia progeny from their more natural polarized epithelial host cells.Chlamydia trachomatis serovars D to K are oculogenital pathogens and the leading cause of bacterial sexually transmitted diseases (41). It is estimated there are 3 to 4 million cases of chlamydial sexually transmitted diseases annually in the United States and some 90 million cases per year worldwide (7). Since the majority of infected individuals are essentially asymptomatic and do not seek medical attention, ascending migration can occur and lead to serious complications, such as prostatitis and epididymitis in men and pelvic inflammatory disease, salpingitis, ectopic pregnancy, and infertility in women (12,14).Chlamydiae are obligate intracellular bacteria and, as such, must be internalized into superficial epithelial cells of the genital mucosa in order to initiate the infectious process. Infection begins with attachment of the infectious elementary bodies (EB) form to the apical surface of columnar epithelial cells, followed by entry via various endocytic mechanisms. The EBcontaining endosomes exit the endocytic pathway to avoid fusion with lysosomes and travel on microtubules to the nuclear hof, where they undergo homotypic fusion with one another, and then the EBs transform into metabolically active reticulate bodies (RB). Since RB divide by binary fission and the number of progeny increases, the expanding endocytic vesicle is termed an inclusion. Eventually, RB mature back into infectious EB, and this devel...

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Characterization of estrogen-responsive epithelial cell lines and their infectivity by genital Chlamydia trachomatis

Cited by 19 publications

References 46 publications

Altered protein secretion of Chlamydia trachomatis in persistently infected human endocervical epithelial cells

Altered protein secretion of Chlamydia trachomatis in persistently infected human endocervical epithelial cells

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

Differences in Chlamydia trachomatis Serovar E Growth Rate in Polarized Endometrial and Endocervical Epithelial Cells Grown in Three-Dimensional Culture

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