scholarly journals Cooperative Interactions between Trichomonas vaginalis and Associated Bacteria Enhance Paracellular Permeability of the Cervicovaginal Epithelium by Dysregulating Tight Junctions

2019 ◽  
Vol 87 (5) ◽  
Author(s):  
Annabel S. Hinderfeld ◽  
Niha Phukan ◽  
Ann-Katrein Bär ◽  
Anthony M. Roberton ◽  
Augusto Simoes-Barbosa
Keyword(s):  
Tight Junction ◽  
Trichomonas Vaginalis ◽  
Bacterial Species ◽  
Cell Monolayer ◽  
Paracellular Permeability ◽  
Vaginal Microbiota ◽  
Gardnerella Vaginalis ◽  
Content Type ◽  
Cooperative Interactions ◽  
Study Results

ABSTRACT The human protozoan Trichomonas vaginalis is the causative agent of trichomoniasis, a prevalent sexually transmitted infection, which is accompanied by a species-diversified vaginal microbiota named community state type IV (CST-IV). Coincidently, CST-IV includes species associated with bacterial vaginosis (e.g. Gardnerella vaginalis, Atopobium vaginae, and Prevotella bivia). Both diseases are linked to the transmission of human immunodeficiency virus (HIV) and preterm birth, which complications are likely to result from the disruption of the cervicovaginal epithelial barrier. Here, we show that paracellular permeability of fluorescein isothiocyanate (FITC)-dextran through a monolayer of human ectocervical cells (hECs) is increased as a consequence of the activity of T. vaginalis and the aforementioned species of CST-IV bacteria cooperatively. T. vaginalis enhances paracellular permeability of hECs two times more than the individual bacterial species, by up to ∼10% versus ∼5%, respectively. However, any two or all three bacterial species are capable of synergizing this effect. T. vaginalis and the bacteria together increase the paracellular permeability of hECs by ∼50%, which is 5 to 10 times more than the results seen with the protozoan or bacteria alone. This effect is accompanied by enhancement of phosphatase activity, while phosphatase inhibition results in preservation of the integrity of the ectocervical cell monolayer. In addition, these microorganisms induce changes in the expression of tight junction proteins, particularly occludin, and of proinflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α). Together, our findings establish that cooperative interactions between CST-IV bacteria and T. vaginalis enhance the paracellular permeability of the cervicovaginal epithelium by disturbing the integrity of the tight junction complex. Our study results highlight the importance of understanding the contribution of the vaginal microbiota to trichomoniasis.

scholarly journals ALactobacillus-Deficient Vaginal Microbiota Dominates Postpartum Women in Rural Malawi

2018 ◽  
Vol 84 (6) ◽  
Author(s):  
Ronan Doyle ◽  
Austridia Gondwe ◽  
Yue-Mei Fan ◽  
Kenneth Maleta ◽  
Per Ashorn ◽  
...  
Keyword(s):  
Bacterial Community ◽  
Birth Outcomes ◽  
Bacterial Species ◽  
Disease Status ◽  
Vaginal Microbiota ◽  
Gardnerella Vaginalis ◽  
Adverse Birth Outcomes ◽  
Content Type ◽  
Inflammatory Conditions ◽  
State Type

ABSTRACTThe bacterial community found in the vagina is an important determinant of a woman's health and disease status. A healthy vaginal microbiota is associated with low species richness and a high proportion of one of a number of differentLactobacillusspp. When disrupted, the resulting abnormal vaginal microbiota is associated with a number of disease states and poor pregnancy outcomes. Studies up until now have concentrated on relatively small numbers of American and European populations that may not capture the full complexity of the community or adequately predict what constitutes a healthy microbiota in all populations. In this study, we sampled and characterized the vaginal microbiota found on vaginal swabs taken postpartum from a cohort of 1,107 women in rural Malawi. We found a population dominated byGardnerella vaginalisand devoid of the most common vaginalLactobacillusspecies, even if the vagina was sampled over a year postpartum. ThisLactobacillus-deficient anaerobic community, commonly labeled community state type (CST) 4, could be subdivided into four further communities. ALactobacillus iners-dominated vaginal microbiota became more common the longer after delivery the vagina was sampled, butG. vaginalisremained the dominant organism. These results outline the difficulty in all-encompassing definitions of what a healthy or abnormal postpartum vaginal microbiota is. Previous identification of community state types and associations among bacterial species, bacterial vaginosis, and adverse birth outcomes may not represent the complex heterogeneity of the microbiota present. (This study has been registered at ClinicalTrials.gov as NCT01239693.)IMPORTANCEA bacterial community in the vaginal tract is dominated by a small number ofLactobacillusspecies, and when not present there is an increased incidence of inflammatory conditions and adverse birth outcomes. A switch to a vaginal bacterial community lacking inLactobacillusspecies is common after pregnancy. In this study, we characterized the postpartum vaginal bacterial community of a large group of women from a resource-poor, undersampled population in rural Malawi. The majority of women were found to have aLactobacillus-deficient community, and even when sampled a year after delivery the majority of women still did not haveLactobacilluspresent in their vaginal microbiota. The effect of becoming pregnant again for those who do not revert to aLactobacillus-dominant community is unknown, and this could suggest that not allLactobacillus-deficient community structures are adverse. A better understanding of this complex community state type is needed.

scholarly journals Cross-Sectional Analysis of Selected Genital Tract Immunological Markers and Molecular Vaginal Microbiota in Sub-Saharan African Women, with Relevance to HIV Risk and Prevention

2015 ◽  
Vol 22 (5) ◽  
pp. 526-538 ◽  
Author(s):  
Jordan K. Kyongo ◽  
Tania Crucitti ◽  
Joris Menten ◽  
Liselotte Hardy ◽  
Piet Cools ◽  
...  
Keyword(s):  
Proinflammatory Cytokines ◽  
Genital Tract ◽  
Bacterial Species ◽  
Vaginal Microbiota ◽  
African Women ◽  
Sexually Active ◽  
Cross Sectional ◽  
Content Type ◽  
Immune Mediators ◽  
Sub Saharan

ABSTRACTData on immune mediators in the genital tract and the factors that modulate them in sub-Saharan women are limited. Cervicovaginal lavage (CVL) samples from 430 sexually active women from Kenya, South Africa, and Rwanda were analyzed for 12 soluble immune mediators using Bio-Plex and Meso Scale Discovery multiplex platforms, as well as single enzyme-linked immunosorbent assays. Ten bacterial species were quantified in vaginal swab samples. Bacterial vaginosis (BV) was defined by Nugent scoring. CVL samples from HIV-infected women showed a clear-cut proinflammatory profile. Pregnant women, adolescents, and women engaging in traditional vaginal practices differed in specific soluble markers compared to reference groups of adult HIV-negative women. Cervical mucus, cervical ectopy, abnormal vaginal discharge, and having multiple sex partners were each associated with an increase in inflammatory mediators. The levels of interleukin-1α (IL-1α), IL-1β, IL-6, IL-12(p70), and IL-8 were elevated, whereas the IL-1RA/IL-1(α+β) ratio decreased in women with BV. The level of gamma interferon-induced protein 10 was lower in BV-positive than in BV-negative women, suggesting its suppression as a potential immune evasion mechanism by BV-associated bacteria.Lactobacillus crispatusandLactobacillus vaginaliswere associated with decreased proinflammatory cytokines and each BV-associated species with increased proinflammatory cytokines. Remarkably, thein vitroanti-HIV activity of CVL samples from BV-positive women was stronger than that of BV-negative women. In conclusion, we found significant associations of factors, including vaginal microbiota, which can influence immune mediators in the vaginal environment in sexually active women. These factors need to be considered when establishing normative levels or pathogenic cutoffs of biomarkers of inflammation and associated risks in African women.

scholarly journals Vaginal Microbiota of the Sexually Transmitted Infections Caused by Chlamydia trachomatis and Trichomonas vaginalis in Women with Vaginitis in Taiwan

2021 ◽  
Vol 9 (9) ◽  
pp. 1864
Author(s):  
Shu-Fang Chiu ◽  
Po-Jung Huang ◽  
Wei-Hung Cheng ◽  
Ching-Yun Huang ◽  
Lichieh Julie Chu ◽  
...  
Keyword(s):  
Chlamydia Trachomatis ◽  
Sexually Transmitted Infections ◽  
Trichomonas Vaginalis ◽  
Vaginal Microbiota ◽  
Gardnerella Vaginalis ◽  
Associated Bacteria ◽  
Sexually Transmitted ◽  
Prevotella Bivia ◽  
Initial Interaction

The three most common sexually transmitted infections (STIs) are Chlamydia trachomatis (CT), Neisseria gonorrhoeae (GC) and Trichomonas vaginalis (TV). The prevalence of these STIs in Taiwan remains largely unknown and the risk of STI acquisition affected by the vaginal microbiota is also elusive. In this study, a total of 327 vaginal swabs collected from women with vaginitis were analyzed to determine the presence of STIs and the associated microorganisms by using the BD Max CT/GC/TV molecular assay, microbial cultures, and 16S rRNA sequencing. The prevalence of CT, TV, and GC was 10.8%, 2.2% and 0.6%, respectively. A culture-dependent method identified that Escherichia coli and Streptococcus agalactiae (GBS) were more likely to be associated with CT and TV infections. In CT-positive patients, the vaginal microbiota was dominated by L. iners, and the relative abundance of Gardnerella vaginalis (12.46%) was also higher than that in TV-positive patients and the non-STIs group. However, Lactobacillus spp. was significantly lower in TV-positive patients, while GBS (10.11%), Prevotella bivia (6.19%), Sneathia sanguinegens (12.75%), and Gemella asaccharolytica (5.31%) were significantly enriched. Using an in vitro co-culture assay, we demonstrated that the growth of L. iners was suppressed in the initial interaction with TV, but it may adapt and survive after longer exposure to TV. Additionally, it is noteworthy that TV was able to promote GBS growth. Our study highlights the vaginal microbiota composition associated with the common STIs and the crosstalk between TV and the associated bacteria, paving the way for future development of health interventions targeting the specific vaginal bacterial taxa to reduce the risk of common STIs.

scholarly journals Specific Vaginal Bacteria Are Associated With an Increased Risk of Trichomonas vaginalis Acquisition in Women

2019 ◽  
Vol 220 (9) ◽  
pp. 1503-1510 ◽  
Author(s):  
Olamide D Jarrett ◽  
Sujatha Srinivasan ◽  
Barbra A Richardson ◽  
Tina Fiedler ◽  
Jacqueline M Wallis ◽  
...  
Keyword(s):  
Ribosomal Rna ◽  
Trichomonas Vaginalis ◽  
Bacterial Species ◽  
Polymerase Chain Reaction Analysis ◽  
Vaginal Microbiota ◽  
Menstrual Phase ◽  
Chain Reaction ◽  
Increased Risk ◽  
16S Ribosomal Rna Gene ◽  
Polymerase Chain

Abstract Background While bacterial vaginosis has been associated with an increased risk of Trichomonas vaginalis (TV) acquisition, it is unknown whether other characteristics of the vaginal microbiota, including the presence of key bacterial species, influence a woman’s risk of TV acquisition. Methods The vaginal microbiota before 25 unique episodes of TV infection involving 18 women was compared to that of 50 controls who remained uninfected. TV was detected by transcription-mediated amplification. Vaginal microbiota were quantified using broad-range polymerase chain reaction analysis and taxon-specific quantitative PCR of the 16S ribosomal RNA gene. Results TV acquisition was significantly associated with the presence of Prevotella amnii (risk ratio [RR], 2.21; 95% confidence interval [CI], 1.12–4.38; P = .02) and Sneathia sanguinegens (RR, 2.58; 95% CI, 1.00–6.62; P = .049). When adjusted for menstrual phase, the association between P. amnii and TV acquisition remained similar (adjusted RR, 2.11; 95% CI, 1.03–4.33; P = .04), but the association between S. sanguinegens and TV acquisition was attenuated (adjusted RR, 2.31; 95% CI, .86–6.23; P = .10). Conclusions Key vaginal bacterial species may contribute to the susceptibility to TV acquisition. Understanding how these bacterial species increase a woman’s risk of TV acquisition could help to guide the development of novel strategies to reduce women’s risk of TV infection.

scholarly journals The ProtozoanTrichomonas vaginalisTargets Bacteria with Laterally Acquired NlpC/P60 Peptidoglycan Hydrolases

mBio ◽  
2018 ◽  
Vol 9 (6) ◽  
Author(s):  
Jully Pinheiro ◽  
Jacob Biboy ◽  
Waldemar Vollmer ◽  
Robert P. Hirt ◽  
Jeremy R. Keown ◽  
...  
Keyword(s):  
Trichomonas Vaginalis ◽  
Close Association ◽  
Transmitted Disease ◽  
Mixed Cultures ◽  
Vaginal Mucosa ◽  
Transmitted Infection ◽  
Content Type ◽  
Sexually Transmitted ◽  
Study Results ◽  
Peptidoglycan Hydrolases

ABSTRACTThe human eukaryotic pathogenTrichomonas vaginaliscauses trichomoniasis, a prevalent sexually transmitted infection. This extracellular protozoan is intimately associated with the human vaginal mucosa and microbiota, but key aspects of the complex interactions between the parasite and the vaginal bacteria remain elusive. We report thatT. vaginalishas acquired, by lateral gene transfer from bacteria, genes encoding peptidoglycan hydrolases of the NlpC/P60 family. Two of theT. vaginalisenzymes were active against bacterial peptidoglycan, retaining the active-site fold and specificity asdl-endopeptidases. The endogenous NlpC/P60 genes are transcriptionally upregulated inT. vaginalisin the presence of bacteria. The overexpression of an exogenous copy enables the parasite to outcompete bacteria from mixed cultures, consistent with the biochemical activity of the enzyme. Our study results highlight the relevance of the interactions of this eukaryotic pathogen with bacteria, a poorly understood aspect of the biology of this important human parasite.IMPORTANCETrichomonas vaginalisis a parasitic protozoan of the human urogenital tract that causes trichomoniasis, a very common sexually transmitted disease. Despite residing extracellularly and in close association with the vaginal bacteria (i.e., the microbiota), very little is known about the nature of the parasite-bacterium interactions. Our study showed that this parasite had acquired genes from bacteria which retained their original function. They produce active enzymes capable of degrading peptidoglycan, a unique polymer of the bacterial cell envelope, helping the parasite to outcompete bacteria in mixed cultures. This study was the first to show that a laterally acquired group of genes enables a eukaryotic mucosal pathogen to control bacterial population. We highlight the importance of understanding the interactions between pathogens and microbiota, as the outcomes of these interactions are increasingly understood to have important implications on health and disease.

scholarly journals Metatranscriptome Analysis of the Vaginal Microbiota Reveals Potential Mechanisms for Protection against Metronidazole in Bacterial Vaginosis

mSphere ◽  
2018 ◽  
Vol 3 (3) ◽  
Author(s):  
Zhi-Luo Deng ◽  
Cornelia Gottschick ◽  
Sabin Bhuju ◽  
Clarissa Masur ◽  
Christoph Abels ◽  
...  
Keyword(s):  
Antibiotic Therapy ◽  
Bacterial Vaginosis ◽  
Bactericidal Effect ◽  
Vaginal Microbiota ◽  
Gardnerella Vaginalis ◽  
Vaginal Fluid ◽  
Content Type ◽  
Production Of Hydrogen ◽  
Cas Genes

ABSTRACT Bacterial vaginosis (BV) is a prevalent multifactorial disease of women in their reproductive years characterized by a shift from the Lactobacillus species-dominated microbial community toward a taxonomically diverse anaerobic community. For unknown reasons, some women do not respond to therapy. In our recent clinical study, among 37 women diagnosed with BV, 31 were successfully treated with metronidazole, while 6 still had BV after treatment. To discover possible reasons for the lack of response in those patients, we performed a metatranscriptome analysis of their vaginal microbiota, comparing them to the patients who responded. Seven of 8 clustered regularly interspaced short palindromic repeat (CRISPR)-associated (Cas) genes of Gardnerella vaginalis were highly upregulated in nonresponding patients. Cas genes, in addition to protecting against phages, might be involved in DNA repair, thus mitigating the bactericidal effect of DNA-damaging agents such as metronidazole. In the second part of our study, we analyzed the vaginal metatranscriptomes of four patients over 3 months and showed high in vivo expression of genes for pore-forming toxins in L. iners and of genes encoding enzymes for the production of hydrogen peroxide and d-lactate in L. crispatus. IMPORTANCE Bacterial vaginosis is a serious issue for women in their reproductive years. Although it can usually be cured by antibiotics, the recurrence rate is very high, and some women do not respond to antibiotic therapy. The reasons for that are not known. Therefore, we undertook a study to detect the activity of the complete microbiota in the vaginal fluid of women who responded to antibiotic therapy and compared it to the activity of the microbiota in women who did not respond. We found that one of the most important pathogens in bacterial vaginosis, Gardnerella vaginalis, has activated genes that can repair the DNA damage caused by the antibiotic in those women that do not respond to therapy. Suppressing these genes might be a possibility to improve the antibiotic therapy of bacterial vaginosis.

scholarly journals Genome Investigation of Urinary Gardnerella Strains and Their Relationship to Isolates of the Vaginal Microbiota

mSphere ◽  
2021 ◽  
Vol 6 (3) ◽  
Author(s):  
Catherine Putonti ◽  
Krystal Thomas-White ◽  
Elias Crum ◽  
Evann E. Hilt ◽  
Travis K. Price ◽  
...  
Keyword(s):  
Urinary Tract ◽  
Lower Urinary Tract Symptoms ◽  
Lower Urinary Tract ◽  
Vaginal Microbiota ◽  
Gardnerella Vaginalis ◽  
Content Type ◽  
Urinary Tract Symptoms ◽  
Species Groups ◽  
Urinary Microbiota ◽  
Lower Urinary

ABSTRACT Gardnerella is a frequent member of the urogenital microbiota. Given the association between Gardnerella vaginalis and bacterial vaginosis (BV), significant efforts have been focused on characterizing this species in the vaginal microbiota. However, Gardnerella also is a frequent member of the urinary microbiota. In an effort to characterize the bacterial species of the urinary microbiota, we present here 10 genomes of urinary Gardnerella isolates from women with and without lower urinary tract symptoms. These genomes complement those of 22 urinary Gardnerella strains previously isolated and sequenced by our team. We included these genomes in a comparative genome analysis of all publicly available Gardnerella genomes, which include 33 urinary isolates, 78 vaginal isolates, and 2 other isolates. While once this genus was thought to consist of a single species, recent comparative genome analyses have revealed 3 new species and an additional 9 groups within Gardnerella. Based upon our analysis, we suggest a new group for the species. We also find that distinction between these Gardnerella species/groups is possible only when considering the core or whole-genome sequence, as neither the sialidase nor vaginolysin genes are sufficient for distinguishing between species/groups despite their clinical importance. In contrast to the vaginal microbiota, we found that only five Gardnerella species/groups have been detected within the lower urinary tract. Although we found no association between a particular Gardnerella species/group(s) and urinary symptoms, further sequencing of urinary Gardnerella isolates is needed for both comprehensive taxonomic characterization and etiological classification of Gardnerella in the urinary tract. IMPORTANCE Prior research into the bacterium Gardnerella vaginalis has largely focused on its association with bacterial vaginosis (BV). However, G. vaginalis is also frequently found within the urinary microbiota of women with and without lower urinary tract symptoms as well as individuals with chronic kidney disease, interstitial cystitis, and BV. This prompted our investigation into Gardnerella from the urinary microbiota and all publicly available Gardnerella genomes from the urogenital tract. Our work suggests that while some Gardnerella species can survive in both the urinary tract and vagina, others likely cannot. This study provides the foundation for future studies of Gardnerella within the urinary tract and its possible contribution to lower urinary tract symptoms.

scholarly journals Vaginolysin Drives Epithelial Ultrastructural Responses to Gardnerella vaginalis

2013 ◽  
Vol 81 (12) ◽  
pp. 4544-4550 ◽  
Author(s):  
Tara M. Randis ◽  
Joanne Zaklama ◽  
Timothy J. LaRocca ◽  
Ferdinand C. O. Los ◽  
Emma L. Lewis ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Bacterial Species ◽  
Gardnerella Vaginalis ◽  
Target Cells ◽  
Vaginal Mucosa ◽  
Content Type ◽  
Induced Membrane ◽  
Membrane Blebbing ◽  
Bleb Formation ◽  
Complex Signaling

ABSTRACTGardnerella vaginalis, the bacterial species most frequently isolated from women with bacterial vaginosis (BV), produces a cholesterol-dependent cytolysin (CDC), vaginolysin (VLY). At sublytic concentrations, CDCs may initiate complex signaling cascades crucial to target cell survival. Using live-cell imaging, we observed the rapid formation of large membrane blebs in human vaginal and cervical epithelial cells (VK2 and HeLa cells) exposed to recombinant VLY toxin and to cell-free supernatants from growing liquid cultures ofG. vaginalis. Binding of VLY to its human-specific receptor (hCD59) is required for bleb formation, as antibody inhibition of either toxin or hCD59 abrogates this response, and transfection of nonhuman cells (CHO-K1) with hCD59 renders them susceptible to toxin-induced membrane blebbing. Disruption of the pore formation process (by exposure to pore-deficient toxoids or pretreatment of cells with methyl-β-cyclodextrin) or osmotic protection of target cells inhibits VLY-induced membrane blebbing. These results indicate that the formation of functional pores drives the observed ultrastructural rearrangements. Rapid bleb formation may represent a conserved response of epithelial cells to sublytic quantities of pore-forming toxins, and VLY-induced epithelial cell membrane blebbing in the vaginal mucosa may play a role in the pathogenesis of BV.

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