Cervicovaginal Inflammation Research Articles

Inflammatory cytokines and a diverse cervicovaginal microbiota associate with cervical dysplasia in a cohort of Hispanics living in Puerto Rico.

Cervical cancer (CC) is women's fourth most common cancer worldwide. A worrying increase in CC rates in Hispanics suggests that besides Human papillomavirus infections, there may be other cofactors included in the epithelial microenvironment that could play a role in promoting the disease. We hypothesized that the cervical microbiome and the epithelial microenvironment favoring inflammation is conducive to disease progression in a group of Hispanics attending gynecology clinics in Puerto Rico. Few studies have focused on the joint microbiota and cytokine profile response in Hispanics outside the US, especially regarding the development of precancerous lesions. We aimed to investigate the relationship between the cervicovaginal microbiome and inflammation in Hispanic women living in PR while considering cervical dysplasia and HPV genotype risk. Cervical samples collected from 91 participants coming to gynecology clinics in San Juan, underwent 16S rRNA genes (V4 region) profiling, and cytokines were measured using Luminex MAGPIX technology. Cytokines were grouped as inflammatory (IL-1β, TNFα, IFNγ, IL-6), anti-inflammatory (IL- 4, IL-10, TGFβ1), and traffic-associated (IL-8, MIP1a, MCP1, IP10). They were related to microbes via an inflammation scoring index based on the quartile and tercile distribution of the cytokine's concentration. We found significant differences in the diversity and composition of the microbiota according to HPV type according to carcinogenic risk, cervical disease, and cytokine abundance. Community State Types (CSTs) represents a profile of microbial communities observed within the vaginal microbiome ecological niche, and Lactobacillus-depleted CST IV had ~ 90% dominance in participants with high-grade squamous intraepithelial lesions and high-risk HPV. The increasing concentration of pro-inflammatory cytokines was associated with a decrease in L. crispatus. In contrast, dysbiosis-associated bacteria such as Gardnerella, Prevotella, Atopobium concomitantly increased with pro-inflammatory cytokines. Our study highlights that the cervical microbiota of Hispanics living in Puerto Rico is composed mostly of diverse CST profiles with decreased Lactobacillus and is associated with a higher pro-inflammatory environment. The joint host-microbe interaction analyses via cytokine and microbiota profiling have very good translational potential.

The role of cervical neutrophils in cervicovaginal inflammation in women at high-risk of delivering preterm

The role of cervical neutrophils in cervicovaginal inflammation in women at high-risk of delivering preterm

Collagen-Based Ovule Therapy Reduces Inflammation and Improve Cervical Epithelialization in Patients with Fungal, Viral, and Bacterial Cervico-Vaginitis.

Background and Objectives: Vulvovaginal infections pose significant health challenges for women, necessitating effective treatment approaches. This retrospective observational study aimed at investigating the efficacy of collagen-based vaginal ovules therapy, specifically Plurigin Ovules, in restoring cervical epithelialization and reducing inflammation in mycotic, viral, and bacterial cervico-vaginitis. Materials and Methods: A total of 398 women with cervico-vaginitis were included in the study, categorized into three groups: bacterial cervico-vaginitis (Group A), viral cervico-vaginitis (Group B), and fungal cervico-vaginitis (Group C). Participants received vaginal therapy with Plurigin Ovules for three months. Vaginal health parameters were assessed at baseline (T0) and after six months (T1) using various diagnostic tests and analyzed with appropriate statistical tests. Results: Significant improvement in cervico-vaginitis was observed in all three groups. At T1, 87.7% patients of Group A, 66.7% of Group B, and 71.5% of Group C achieved infection resolution (all p < 0.05). Positive colposcopy results decreased across all groups (p < 0.001). Positive vaginal swabs and altered vaginal pH decreased in group A and C (p < 0.001). Positive HPV tests decreased in Group B (p < 0.001). Positive Pap tests and clinical examinations decreased significantly across all groups (p < 0.001). The odds ratios were calculated to reveal the significant associations between these diagnostic outcomes. The therapy was well-tolerated, and no major adverse events were reported. Conclusion: Plurigin Ovules exhibited promising therapeutic outcomes in the three cervico-vaginitis conditions studied. Bacterial cervico-vaginitis showed the most significant improvement, followed by fungal and viral cervico-vaginitis. These findings emphasize the potential of Plurigin Ovules as an effective therapeutic option for cervico-vaginal inflammation and infection, highlighting its role in promoting re-epithelialization and reducing inflammatory processes in the cervix and vagina.

Increased genital mucosal cytokines in Canadian women associate with higher antigen-presenting cells, inflammatory metabolites, epithelial barrier disruption, and the depletion of L. crispatus

BackgroundCervicovaginal inflammation has been linked to negative reproductive health outcomes including the acquisition of HIV, other sexually transmitted infections, and cervical carcinogenesis. While changes to the vaginal microbiome have been linked to genital inflammation, the molecular relationships between the functional components of the microbiome with cervical immunology in the reproductive tract are understudied, limiting our understanding of mucosal biology that may be important for reproductive health.ResultsIn this study, we used a multi’-omics approach to profile cervicovaginal samples collected from 43 Canadian women to characterize host, immune, functional microbiome, and metabolome features of cervicovaginal inflammation. We demonstrate that inflammation is associated with lower amounts of L. crispatus and higher levels of cervical antigen-presenting cells (APCs). Proteomic analysis showed an upregulation of pathways related to neutrophil degranulation, complement, and leukocyte migration, with lower levels of cornified envelope and cell-cell adherens junctions. Functional microbiome analysis showed reductions in carbohydrate metabolism and lactic acid, with increases in xanthine and other metabolites. Bayesian network analysis linked L. crispatus with glycolytic and nucleotide metabolism, succinate and xanthine, and epithelial proteins SCEL and IVL as major molecular features associated with pro-inflammatory cytokines and increased APCs.ConclusionsThis study identified key molecular and immunological relationships with cervicovaginal inflammation, including higher APCs, bacterial metabolism, and proteome alterations that underlie inflammation. As APCs are involved in HIV transmission, parturition, and cervical cancer progression, further studies are needed to explore the interactions between these cells, bacterial metabolism, mucosal immunity, and their relationship to reproductive health.BGqtALm3-LNazbdZNFi1WRVideo

Does Lactobacillus Exert a Protective Effect on the Development of Cervical and Endometrial Cancer in Women?

Simple SummaryCervical cancer is the fourth most common cancer in women worldwide. Tumour-related deaths are most frequent in low- and middle-income countries. Currently, most vaccines against human papillomavirus (HPV) are based on virus-like particles; they protect against HPV infection but have no therapeutic effects. Because dysbiosis has been shown to increase cancer risk, lactic acid bacteria (LAB)-based vaccines, which have been shown to have an immunomodulatory effect, have recently attracted attention. Mucosal immunization with viable colonies of Lactobacillus via intranasal, intravaginal, and oral routes to decrease the risk of cervical cancer seems promising; thus, such research is of high value. While advances have been made in understanding associations between microbiota dysregulation and carcinogenesis, further studies are required to identify the underlying cellular mechanisms and to confirm previous findings. This manuscript summarizes available data concerning the impact of microbiota on cancer risk and presents recent strategies to fight cervical and endometrial cancers.Cervical cancer is a significant health problem with increasing occurrence and mortality. This infection-associated tumour is caused by the human papillomavirus (HPV). HPV infection is cleared by the immune system within 6–18 months in most patients; however, persistent high-risk HPV (hrHPV) infections can lead to the development of cervical cancer. Virus persistence is promoted by immunodeficiency, Chlamydia trachomatis infection, smoking, and age, as well as the imbalance of cervicovaginal microbiota and inflammation. The abundance of bacteria in the vagina favours the maintenance of a dynamic balance; their coexistence influences health or disease states. The eubiotic vaginal microbiota of reproductive-aged women is composed mostly of various Lactobacillus species (spp.), which exert protective effects via the production of lactic acid, bacteriocins, polysaccharides, peptidoglycans, and hydrogen peroxide (H2O2), lowering pH, raising the viscosity of cervicovaginal mucus, and hampering both the adhesion of cells to epithelial tissue and the entry of HPV. The depletion of beneficial microorganisms could increase the risk of sexually transmitted infections. Emerging therapies involve mucosal, intranasal vaccines, which trigger systemic and mucosal immune responses, thus protecting against HPV-induced tumours. The use of probiotics has also been suggested to affect various biological processes associated with tumourigenesis (inflammation, oxidative stress, apoptosis, proliferation, and metastasis).

Abstract 712: Cytokine and TGF-β levels are associated with changes in cervicovaginal microbiota in a cohort of Caribbean women

Abstract Recent studies suggest that the cervical microbiome can strongly influence inflammation and pre-cancerous lesion progression. However, research focused on understanding the role of microbial communities in the progression of pre-cancerous lesions to cancer in Latino women is scarce. We aimed to investigate the relationship between the cervicovaginal microbiome and inflammation while considering cervical neoplasia and HPV infection in Puerto Rican women. We collected cervical swabs and lavages from 142 participants coming to colposcopy clinics in San Juan, Puerto Rico. Genomic DNA was extracted from swabs, and 16S rDNA V4 region genes were amplified and sequenced by Illumina MiSeq. Inflammatory (IL-1β, TNFa, IFNg, IL-6), anti-inflammatory (IL-4, IL-10, TGFβ1), and trafficking (IL-8, MIP1a, MCP1, IP10) cytokines were measured from cervical lavages, using Luminex MAGPIX technology. Cytokines were related to microbes via an inflammation scoring index based on the quartile and tercile distribution of the cytokine’s concentration. IL-10 (p value= 0.0455) was significantly different when evaluating HPV risk, while IL1-β (p value= 0.0005) and INF-γ (p value= 0.0258) were significant when evaluating cervical disease. We found significant differences in diversity and composition of the microbiota among HPV risk, cervical disease, pro-inflammatory, anti-inflammatory, and trafficking cytokine abundances. The increasing concentration of IL1-β, IL-10, and INF-γ, associated with a decrease in Lactobacillus communities. While contrarily, bacteria associated with dysbiosis such as Gardnerella, Prevotella, Atopobium increased. This study also revealed that the most dominant community state types (CST) among Puerto Rican women regardless of lesion or HPV status, are CST3 and 4 featuring high diversity and anaerobic bacteria typical of vaginosis in Caucasians. These CSTs are especially abundant with ~ 90% dominance in participants with high grade disease (HGSIL) and high-risk HPV. Our study evidence that the cervical microbiota of Puerto Ricans is characteristically diverse and that the joint host-microbe interaction analyses via cytokine signaling and microbiota in pre-cancerous lesions has great translational potential. Citation Format: Eduardo Lemuel Tosado Rodríguez, Anelisse Dominicci-Maura, Loyda Mendez, Stephanie Dorta, Josefina Romaguera, Filipa Godoy-Vitorino. Cytokine and TGF-β levels are associated with changes in cervicovaginal microbiota in a cohort of Caribbean women [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 712.

Considerations for Choosing Soluble Immune Markers to Determine Safety of Novel Vaginal Products.

Several soluble cytokines have been associated with microbicide-induced cervicovaginal inflammation, non-optimal vaginal microbiota, and risk of HIV acquisition. Many of these biomarkers are used in preclinical assays to estimate the safety of vaginally applied products. However, there are currently no validated biomarkers to evaluate the safety of novel vaginal products in clinical trials. This hinders the rapid and rational selection of novel products being tested in first-in-human trials. We reviewed available literature to assess how best to select and measure soluble immune markers to determine product safety in first in human clinical trials of novel vaginal products.

From Microbiome to Inflammation: The Key Drivers of Cervical Cancer.

Cervical cancer is the third leading cause of cancer-related death worldwide. Microbes and hosts form a mutually beneficial symbiosis relationship, and various parts of the host body are microbial habitats. Microbes can trigger inflammation in certain parts of the host body, contributing to cervical cancer development. This article reviews the relationship between cervicovaginal microbes, inflammation and cervical cancer, and discusses the effect of some key cervical microbes on cervical cancer. Finally, probiotic therapy and immunotherapy are summarized.

Distinct Immune Responses Elicited From Cervicovaginal Epithelial Cells by Lactic Acid and Short Chain Fatty Acids Associated With Optimal and Non-optimal Vaginal Microbiota.

Non-optimal vaginal microbiota, as observed in bacterial vaginosis (BV), is typically characterized by a depletion of beneficial lactobacilli and an abundance of numerous anaerobes. These non-optimal conditions are associated with subclinical cervicovaginal inflammation and an increased risk of HIV infection compared to women colonized with optimal vaginal microbiota dominated by lactobacilli. Lactic acid (LA) is a major organic acid metabolite produced by vaginal lactobacilli that elicits anti-inflammatory effects from cervicovaginal epithelial cells and is dramatically depleted during BV. However, it is unclear if LA retains its anti-inflammatory activity in the presence of vaginal microbiota metabolites comprising short chain fatty acids (SCFAs) and succinic acid, which are also produced by an optimal vaginal microbiota. Furthermore, the immunomodulatory effect of SCFAs and succinic acid on cervicovaginal epithelial cells at higher concentrations present during BV is unknown. Here we report that in the presence of physiologically relevant concentrations of SCFAs and succinic acid at pH 3.9 (as found in women with lactobacillus-dominated microbiota) LA induced an anti-inflammatory state in cervicovaginal epithelial cells and inhibited inflammation elicited by the toll-like receptor (TLR) agonists polyinosinic:polycytidylic acid and Pam3CSK4. When cervicovaginal epithelial cells were treated with a vaginal microbiota metabolite mixture representative of BV, containing a lower concentration of LA but higher concentrations of SCFA/succinic acid at pH 7, no anti-inflammatory was observed. Rather, the vaginal microbiota metabolite mixture representative of BV dysregulated the immune response of cervicovaginal epithelial cells during prolonged and sustained treatments. This was evidenced by increased basal and TLR-induced production of pro-inflammatory cytokines including tumor necrosis factor-α, but decreased basal production of chemokines including RANTES and IP-10. Further characterization of individual components of the BV vaginal microbiota mixture suggested that acetic acid is an important vaginal microbiota metabolite capable of eliciting diverse immunomodulatory effects on a range of cervicovaginal epithelial cell targets. These findings indicate that elevated levels of SCFAs are a potential source of cervicovaginal inflammation in women experiencing BV, and support the unique anti-inflammatory properties of LA on cervicovaginal epithelial cells as well as a role for LA or LA-producing lactobacilli to reverse genital inflammation associated with increased HIV risk.

Associations between vaginal bacteria implicated in HIV acquisition risk and proinflammatory cytokines and chemokines

ObjectivesRecent studies have identified vaginal bacterial taxa associated with increased HIV risk. A possible mechanism to explain these results is that individual taxa differentially promote cervicovaginal inflammation. This study aimed...

Impact of Herpes Simplex Virus Type 2 and Human Immunodeficiency Virus Dual Infection on Female Genital Tract Mucosal Immunity and the Vaginal Microbiome.

Mechanisms linking herpes simplex virus type 2 (HSV-2) with human immunodeficiency virus (HIV) are not fully defined. We tested the hypothesis that HSV-2 and HIV dual infection is associated with cervicovaginal inflammation and/or vaginal dysbiosis. Genital tract samples were obtained weekly over a 12-week period from 30 women seropositive (+) for HIV and HSV-2 and 15 women each who were seropositive for one or seronegative (-) for both viruses. Immune mediators, antimicrobial activity, and microbial composition and diversity were compared. Significant differences in the concentrations of interferon-γ (P = .002), tumor necrosis factor-α (P = .03), human beta defensin 1 (P = .001), secretory leukocyte protease inhibitor (P = .01), and lysozyme (P = .03) were observed across the 4 groups (Kruskal-Wallis). There were also significant differences in vaginal microbial alpha diversity (Simpson index) (P = .0046). Specifically, when comparing HIV-1+/HSV-2+ to HIV-1-/HSV-2- women, a decrease in Lactobacillus crispatus and increase in diverse anaerobes was observed. The number of genital HSV outbreaks was greater in HIV+ versus HIV- women (39 versus 12) (P = .04), but there were no significant differences when comparing outbreak to non-outbreak visits. Increased microbial diversity and cervicovaginal inflammation in HIV and HSV-2 dually infected women may adversely impact genital health and, in the absence of antiretroviral therapy, facilitate HIV shedding.

The relationship of cervical microbiota diversity with race and disparities in preterm birth.

Pregnant non-Hispanic blacks (NHB) have increased vaginal microbiome diversity compared to non-Hispanic whites (NHW) which may contribute to increased preterm birth. Cervical microbiome diversity is poorly characterized in pregnancy, therefore our objective was to correlate cervical microbiota diversity with cervico-vaginal inflammation by race and delivery timing. Pregnant women were recruited in the first and second trimesters. A sterile cervical swab and saline lavage were collected at a single time point. Using 16S rRNA sequencing, Chao1 and Shannon Diversity (SDI) indicies were measured and compared by race and delivery timing (preterm vs. term delivery). Cervico-vaginal inflammatory markers were also compared by race and delivery timing. Spearman correlation coefficients between cervical microbiome diversity and cervico-vaginal inflammatory markers were calculated. Of the 51 subjects, 39 (76%) were NHB and 12 (24%) were NHW. Cervical microbiota SDI was significantly higher in NHB compared to NHW (0.5 vs. 0.1; p = 0.03). However, there were no difference in Chao1 diversity or cervico-vaginal inflammatory markers by race or delivery timing. Our findings suggest the cervical microbiota diversity during pregnancy differs by race. Larger cohort studies will further determine if altered cervical diversity is part of the pathogenesis of PTB and explains race disparities.

The vaginal microbiome amplifies sex hormone-associated cyclic changes in cervicovaginal inflammation and epithelial barrier disruption.

Susceptibility to HIV is associated with the menstrual cycle and vaginal microbiome, but their collective impact on vaginal inflammation remains unclear. Here, we characterized the cervicovaginal proteome, inflammation, and microbiome community structure and function during the menstrual cycle. Cervicovaginal secretions were collected from regularly cycling women (n=16) at median day 10, 16, and 24 of each menstrual cycle and analyzed by mass spectrometry, 16S rRNA gene sequencing, and a multiplex bead array immunoassay. Follicular, ovulatory, and luteal phases were defined by serum sex hormone levels. Ovulation showed the largest mucosal proteome changes, where 30% and 19% of the 406 human proteins identified differed compared to the luteal and follicular phases, respectively. Neutrophil/leukocyte migration pathways were lowest during ovulation and peaked in the luteal phase, while antimicrobial and epithelial barrier promoting proteins were highest during ovulation. Vaginal microbial community structure and function did not vary significantly during the menstrual cycle, with the majority consistently Lactobacillus-dominant (63%) or non-Lactobacillus-dominant (25%). Fluctuations in the epithelial barrier protein RPTN between the ovulatory and luteal phase were amplified in women with Gardnerella vaginalis and anaerobic bacteria and reduced when Lactobacillus was dominant. This small study demonstrates that sex hormones modulate neutrophil/leukocyte inflammation, barrier function, and antimicrobial pathways in the female genital tract with the strongest changes occurring during ovulation. The data further suggest a microbiome context for hormone-driven changes in vaginal immunity which may have implications for HIV susceptibility.

Cervicovaginal Inflammation Facilitates Acquisition of Less Infectious HIV Variants.

We evaluated whether genital inflammation affects the selection of the transmitted virus. Among South African women, we found that preinfection genital inflammation facilitates transmission of less infectious human immunodeficiency virus, but highly infectious viruses are able to establish infection regardless of inflammation status. This suggests that viral phenotype can influence transmission risk.

Cervicovaginal microbiome dysbiosis is associated with proteome changes related to alterations of the cervicovaginal mucosal barrier.

Vaginal microbiome (VMB) dysbiosis is associated with increased acquisition of HIV. Cervicovaginal inflammation and other changes to the mucosal barrier are thought to have important roles but human data are scarce. We compared the human cervicovaginal proteome by mass spectrometry of 50 Rwandan female sex workers who had previously been clustered into four VMB groups using a 16S phylogenetic microarray; in order of increasing bacterial diversity: Lactobacillus crispatus-dominated VMB (group 1), Lactobacillus iners-dominated VMB (group 2), moderate dysbiosis (group 3), and severe dysbiosis (group 4). We compared relative protein abundances among these VMB groups using targeted (abundance of pre-defined mucosal barrier proteins) and untargeted (differentially abundant proteins among all human proteins identified) approaches. With increasing bacterial diversity, we found: mucus alterations (increasing mucin 5B and 5AC), cytoskeleton alterations (increasing actin-organizing proteins; decreasing keratins and cornified envelope proteins), increasing lactate dehydrogenase A/B as markers of cell death, increasing proteolytic activity (increasing proteasome core complex proteins/proteases; decreasing antiproteases), altered antimicrobial peptide balance (increasing psoriasin, calprotectin, and histones; decreasing lysozyme and ubiquitin), increasing pro-inflammatory cytokines, and decreasing immunoglobulins immunoglobulin G1/2. Although temporal relationships cannot be derived, our findings support the hypothesis that dysbiosis causes cervicovaginal inflammation and other detrimental changes to the mucosal barrier.

O13.4 Cervicovaginal microbiome dysbiosis is associated with proteome changes related to alterations of the cervicovaginal mucosal barrier

Introduction Vaginal microbiome (VMB) dysbiosis is associated with increased acquisition of HIV and sexually transmitted infections (STIs). Cervicovaginal inflammation and other changes to the mucosal barrier are thought to play important roles but human data are scarce. In this study, we compared the cervicovaginal proteome among women with different VMB compositions. Methods Cervicovaginal lavages of 50 Rwandan female sex workers with known VMB composition were selected for human proteome analysis using mass-spectrometry. These women were previously clustered into four VMB groups in order of increasing bacterial diversity: group 1 had a Lactobacillus crispatus-dominated VMB; group 2 a L. iners -dominated VMB; group 3 moderate dysbiosis; and group 4 severe dysbiosis. We compared relative protein abundances among these VMB groups using targeted (abundance of pre-defined mucosal barrier proteins) and untargeted (differentially abundant proteins among all human proteins identified) approaches. Results With increasing bacterial diversity, we found: mucus alterations (increasing mucin 5B and 5AC), cytoskeleton alterations (increasing actin-organising proteins; decreasing keratins and cornified envelope proteins), increasing cell death (using LDHA/B as biomarkers of cell death), altered proteolytic activity (increasing proteasome core complex proteins/proteases; decreasing antiproteases), altered antimicrobial peptide balance (increasing psoriasin, calprotectin, and histones; decreasing lysozyme and ubiquitin), increasing proinflammatory cytokines, and decreasing immunoglobulins IgG1/2. Conclusion The VMB is strongly associated with the cervicovaginal human proteome in this cohort of Rwandan women at high risk of HIV and other STIs. Although temporal relationships cannot be derived, our findings support the hypothesis that dysbiosis causes cervicovaginal inflammation and other detrimental changes to the mucosal barrier that may lead to increased HIV/STI acquisition. Disclosure of interest statement This work was funded by the Institute of Infection and Global Health of the University of Liverpool, the Aids Fonds Netherlands (project number 201102), European and Developing Countries Clinical Trials Partnership (project number CT.2005.33070.001) and the European Commission (CHAARM consortium). The authors declare no conflicts of interest.

S13.4 The vaginal microbiome and sti

Our systematic review of 63 molecular vaginal microbiome (VMB) studies conducted between 2008 and 2013 confirmed that lactobacilli-dominated VMB are associated with a healthy vaginal micro-environment and that bacterial vaginosis (BV) is best described as a polybacterial dysbiosis. However, not all lactobacilli seem to be equally healthy (e.g. a Lactobacillus iners-dominated VMB is more likely to shift to dysbiosis than an L. crispatus -dominated VMB and L. iners often remains present during dysbiosis) and multiple dysbiosis entities with clinical relevance seem to exist. Some women with BV have a highly diverse VMB consisting of high loads of planktonic anaerobic bacteria, while others have a vaginal biofilm (typically including Gardnerella vaginalis ). Furthermore, a small but clinically relevant proportion of women have a VMB dominated by Streptococci and/or Escherichia coli . Vaginal colonisation with Candida spp. is more common in women with a lactobacilli-dominated VMB than in women with dysbiosis, and women who receive antibiotic treatment for BV often subsequently develop vaginal candidiasis. Research has shown that all of these conditions cause disruption of the cervicovaginal mucosal barrier as well as cervicovaginal inflammation, which in turn might cause serious complications such as increased HIV acquisition, pre-term birth, and maternal/neonatal sepsis. At the moment, asymptomatic BV (by microscopic and clinical criteria) is usually not treated and symptomatic BV is treated with oral or vaginal metronidazole or clindamycine. However, recurrence rates are very high (up to 50% within six months) and novel treatments – such as vaginal biofilm disruption and vaginal probiotics – are therefore needed. Furthermore, the different dysbiosis entities, or combinations of entities, likely require different clinical management approaches. Systems biology has now become more affordable and should be incorporated into epidemiological studies to address associations of different dysbiotic entities with clinical outcomes, and to evaluate interventions aimed at restoring and maintaining a lactobacilli-dominated VMB.

Bacterial vaginosis and the cervicovaginal immune response.

Bacterial vaginosis (BV) is a common cause of vaginal discharge in reproductive age women around the world and is associated with several poor reproductive health outcomes, including HIV-1 acquisition. One possible mechanism for this association is the inflammatory immune response induced by BV in the cervical and vaginal mucosae. There is significant heterogeneity in reports of markers of cervicovaginal inflammation in women with BV, likely due to microbial and host diversity, as well as differences in study design. In this article, we review the characteristics of the mucosal immune response in BV, the potential role of lactobacilli in modulating that response, and the impact of individual BV-associated bacterial species on mucosal immunity. We focus on inflammatory markers that are proposed to increase the risk of HIV-1 acquisition.

In Captive Rhesus Macaques, Cervicovaginal Inflammation Is Common but Not Associated with the Stable Polymicrobial Microbiome

Vaginal inoculation of rhesus macaques (RM) with simian immunodeficiency virus (SIV) has been used to study the biology of HIV transmission. Although the results of vaginal SIV transmission experiments could be affected by vaginal inflammation, studies to date have been conducted without regard to levels of pre-existing genital inflammation present in RM. We collected cevicovaginal secretions (CVS) from 33–36 RM during the mid menstrual cycle (day 10–20) at 2 time points approximately 8 months apart and characterized the mRNA and protein levels of inflammatory cytokines, chemokines and interferon-stimulated genes. There was extreme variability in the levels of inflammatory mediators (IFN-α, IFN-γ, IL-6, TNF, IL-1b, IP-10, MIG, IL-12 and IL-17). In most animals, the mRNA levels of the inflammatory mediators were similar in the 2 CVS samples collected 8 months apart, suggesting that genital inflammation is stable in a subset of captive female RM. At both time points the cervicovaginal microbiota had low levels of Lactobacillus and was relatively diverse with an average of 13 genera in the samples from the first time point (median 13, range 7–21) and an average of 11.5 genera in the samples from the second time point (median 11, range 5–20). Many of the macaques had similar microbiota in the samples collected 8 months apart. However, we found no correlation between specific bacterial genera and the mRNA or protein levels of the inflammatory mediators in the genital tract of RM in this study. It seems likely that results of published vaginal SIV transmission experiments in RM have been influenced by pre-existing inflammation in the animals used for the experiments.

L-Selectin and P-Selectin Are Novel Biomarkers of Cervicovaginal Inflammation for Preclinical Mucosal Safety Assessment of Anti-HIV-1 Microbicide

A major obstacle thwarting preclinical development of microbicides is the lack of a validated biomarker of cervicovaginal inflammation. Therefore, the present study aims to identify novel noninvasive soluble markers in a murine model for assessment of microbicide mucosal safety. By performing cytokine antibody array analysis, we identified two adhesion molecules, L-selectin and P-selectin, which significantly increased when mucosal inflammation was triggered by nonoxynol-9 (N9), an anti-HIV-1 microbicide candidate that failed clinical trials, in a refined murine model of agent-induced cervicovaginal inflammation. We found that patterns of detection of L-selectin and P-selectin were obviously different from those of the two previously defined biomarkers of cervicovaginal inflammation, monocyte chemotactic protein 1 (MCP-1) and interleukin 6 (IL-6). The levels of these two soluble selectins correlated better than those of MCP-1 and IL-6 with the duration and severity of mucosal inflammation triggered by N9 and two approved proinflammatory compounds, benzalkonium chloride (BZK) and sodium dodecyl sulfate (SDS), but not by two nonproinflammatory compounds, carboxymethyl celluose (CMC; microbicide excipients) and tenofovir (TFV; microbicide candidate). These data indicated that L-selectin and P-selectin can serve as additional novel cervicovaginal inflammation biomarkers for preclinical mucosal safety evaluation of candidate microbicides for the prevention of infection with HIV and other sexually transmitted pathogens.