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Effect of 17β-estradiol on a human vaginal Lactobacillus crispatus strain.

ABSTRACT: Lactobacilli and estrogens play essential roles in vaginal homeostasis. We investigated the potential direct effect of 17β-estradiol on a vaginal strain of Lactobacillus crispatus, the major bacterial species of the vaginal microbiota. 17β-estradiol (10^-6 to 10^-10 M) had no effect on L. crispatus growth, but markedly affected the membrane dynamics of this bacterium. This effect appeared consistent with a signal transduction process. The surface polarity and aggregation potential of the bacterium were unaffected by exposure to 17β-estradiol, but its mean size was significantly reduced. 17β-estradiol also promoted biosurfactant production by L. crispatus and adhesion to vaginal VK2/E6E7 cells, but had little effect on bacterial biofilm formation activity. Bioinformatic analysis of L. crispatus identified a membrane lipid raft-associated stomatin/prohibitin/flotillin/HflK domain containing protein as a potential 17β-estradiol binding site. Overall, our results reveal direct effects of 17β-estradiol on L. crispatus. These effects are of potential importance in the physiology of the vaginal environment, through the promotion of lactobacillus adhesion to the mucosa and protection against pathogens.

SUBMITTER: Clabaut M

PROVIDER: S-EPMC8010061 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundLactobacillus crispatus is the dominant species in the vaginal microbiota associated with health and considered a homeostasis biomarker. Interestingly, some strains are even used as probiotics. However, the genetic mechanisms of L. crispatus involved in the control of the vaginal microbiome and protection against bacterial vaginosis (BV) are not entirely known. To further investigate these mechanisms, we sequenced and characterized the first four L. crispatus genomes from vaginal samples from Brazilian women and used genome-wide association study (GWAS) and comparative analyses to identify genetic mechanisms involved in healthy or BV conditions and selective pressures acting in the vaginal microbiome.MethodsThe four genomes were sequenced, assembled using ten different strategies and automatically annotated. The functional characterization was performed by bioinformatics tools comparing with known probiotic strains. Moreover, it was selected one representative strain (L. crispatus CRI4) for in vitro detection of phages by electron microscopy. Evolutionary analysis, including phylogeny, GWAS and positive selection were performed using 46 public genomes strains representing health and BV conditions.ResultsGenes involved in probiotic effects such as lactic acid production, hydrogen peroxide, bacteriocins, and adhesin were identified. Three hemolysins and putrescine production were predicted, although these features are also present in other probiotic strains. The four genomes presented no plasmids, but 14 known families insertion sequences and several prophages were detected. However, none of the mobile genetic elements contained antimicrobial resistance genes. The genomes harbor a CRISPR-Cas subtype II-A system that is probably inactivated due to fragmentation of the genes csn2 and cas9. No genomic feature was associated with a health condition, perhaps due to its multifactorial characteristic. Five genes were identified as under positive selection, but the selective pressure remains to be discovered. In conclusion, the Brazilian strains investigated in this study present potential protective properties, although in vitro and in vivo studies are required to confirm their efficacy and safety to be considered for human use.

Project description:BackgroundThe vaginal microbiota is associated with the health of the female reproductive system and the offspring. Lactobacillus crispatus belongs to one of the most important vaginal probiotics, while its role in the agglutination and immobilization of human sperm, fertility, and offspring health is unclear.MethodsAdherence assays, sperm motility assays, and Ca2+-detecting assays were used to analyze the adherence properties and sperm motility of L. crispatus Lcr-MH175, attenuated Salmonella typhimurium VNP20009, engineered S. typhimurium VNP20009 DNase I, and Escherichia coli O157:H7 in vitro. The rat reproductive model was further developed to study the role of L. crispatus on reproduction and offspring health, using high-throughput sequencing, real-time PCR, and molecular biology techniques.ResultsOur results indicated that L. crispatus, VNP20009, VNP20009 DNase I, and E. coli O157:H7 significantly inhibited the sperm motility in vitro via adversely affecting the sperm intracellular Ca2+ concentration and showed a high adhesion to sperms. The in vivo results indicated that L. crispatus and other tested bacteria greatly reduced the pregnancy rates, but L. crispatus had a positive effect on maternal health and offspring development. Moreover, the transplantation of L. crispatus could sustain a normal bacterial composition of the vaginal microbiota in healthy rats and markedly reduced the expression of uterine inflammatory factors (toll-like receptor-4/nuclear factor kappa-B, tumor necrosis factor-α, production of interleukin-1β, etc.) and apoptosis factors (Fas Ligand, Bcl-2-associated X protein/B cell lymphoma-2, etc.) compared with the other tested strains.ConclusionOur study demonstrated that the vaginal probiotic L. crispatus greatly affected the sperm activity and could also reduce pregnancies through its adhesion property, which might account for some unexplained infertility. Therefore, more caution should be paid when using L. crispatus as a vaginal viable preparation in women of child-bearing age, especially for women whose partners have abnormal sperms.

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Effect of 17β-estradiol on a human vaginal Lactobacillus crispatus strain.

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