molBV reveals immune landscape of bacterial vaginosis and predicts human papillomavirus infection natural history

Bacterial vaginosis (BV) is a highly prevalent condition that is associated with adverse health outcomes. It has been proposed that BV’s role as a pathogenic condition is mediated via bacteria-induced inflammation. However, the complex interplay between vaginal microbes and host immune factors has yet to be clearly elucidated. Here, we develop molBV, a 16 S rRNA gene amplicon-based classification pipeline that generates a molecular score and diagnoses BV with the same accuracy as the current gold standard method (i.e., Nugent score). Using 3 confirmatory cohorts we show that molBV is independent of the 16 S rRNA region and generalizable across populations. We use the score in a cohort without clinical BV states, but with measures of HPV infection history and immune markers, to reveal that BV-associated increases in the IL-1β/IP-10 cytokine ratio directly predicts clearance of incident high-risk HPV infection (HR = 1.86, 95% CI: 1.19-2.9). Furthermore, we identify an alternate inflammatory BV signature characterized by elevated TNF-α/MIP-1β ratio that is prospectively associated with progression of incident infections to CIN2 + (OR = 2.81, 95% CI: 1.62-5.42). Thus, BV is a heterogeneous condition that activates different arms of the immune response, which in turn are independent risk factors for HR-HPV clearance and progression. Clinical Trial registration number: The CVT trial has been registered under: NCT00128661.

Cervicovaginal Microbiome Factors in Clearance of Human Papillomavirus Infection

Persistent high-risk human papillomavirus (hrHPV) infection is the highest risk to cervical cancer which is the fourth most common cancer in women worldwide. A growing body of literatures demonstrate the role of cervicovaginal microbiome (CVM) in hrHPV susceptibility and clearance, suggesting the promise of CVM-targeted interventions in protecting against or eliminating HPV infection. Nevertheless, the CVM-HPV-host interactions are largely unknown. In this review, we summarize imbalanced CVM in HPV-positive women, with or without cervical diseases, and the progress of exploring CVM resources in HPV clearance. In addition, microbe- and host-microbe interactions in HPV infection and elimination are reviewed to understand the role of CVM in remission of HPV infection. Lastly, the feasibility of CVM-modulated and -derived products in promoting HPV clearance is discussed. Information in this article will provide valuable reference for researchers interested in cervical cancer prevention and therapy.

The Immune Landscape of Molecular Bacterial Vaginosis and HPV Natural History

Bacterial vaginosis (BV) is a highly prevalent condition that is associated with acquisition of sexually transmitted infections and adverse reproductive outcomes. It has been proposed that BV’s role as a pathogenic condition is mediated via bacteria-induced local inflammation. However, the complex interplay between vaginal microbes and host immune factors has yet to be clearly elucidated. We demonstrate that 16S rRNA amplicon sequencing and a novel pipeline can be used to generate a molecular Nugent BV score (molBV) corresponding to the Nugent score 0 - 10. This algorithm is independent of the region of 16S rRNA amplified, the sequencing platform and source population. We further identify two local immune cytokine patterns associated with this molecular Nugent score (q-values<0.001). The main immune response is represented by an elevated IL-1β/IP-10 ratio, whereas a second pattern consists of an increased TNF-α/MIP-1β ratio. To evaluate the biological significance of molBV-BV and the local immune response, we show that clearance of high-risk HPV (HR-HPV) infections (HZ=1.86, 95% CI: 1.19-2.9) was associated with immune profiles, but not molBV-BV when both were considered in the model. In contrast, the TNF-α/MIP-1β signature was associated with progression of incident infections to CIN2+ (OR = 2.81, 95% CI: 1.62-5.42), but not HR-HPV clearance. Thus, BV is a heterogeneous condition that activates different arms of the local immune response, which in turn are independent risk factors for HR-HPV clearance and progression.

Treatment with an intravaginal gel containing siliceous dioxide, selenite, and citric acid to promote regression of ASC-US, LSIL, ASC-H, HSIL, p16/Ki67 status and improve clearance of hr-HPV in cervical specimens

Objective: An intravaginal gel containing highly dispersed silicon dioxide (SiO2) and an anti-oxidative combination of citric acid and sodium selenite was tested for its ability to promote regression of abnormal cytological findings and its influence on hr-HPV status and tumor markers p16/Ki67 (CINtec® PLUS test).Patients and methods: A control study was performed, including women (n=100) diagnosed with conspicuous cervical smears (ASC-US, LSIL, ASC-H, or HSIL). The gel was applied for 3x28 days. After three months, participants were analyzed for Pap status, hr-HPV strains, and expression of tumor markers p16/Ki67. Three months later, Pap testing and p16/Ki67 analysis were repeated. The results were compared to those of 106 women who met the same inclusion criteria but did not obtain any treatment.Results: After six months, cytological Pap findings were improved in 80.9% of the participants in the treated group, and the clearing of hr-HPV was observed in 53% of cases. Only 5.3% were tested p16/Ki67 positive after six months in comparison to 75.0% at baseline. In the comparison group, 37.1% of the Pap smears and 18.6% of the CinTec results were improved, but no hr-HPV clearance was observed. The improvements were highly significant for the treatment. Conclutions: The vaginal gel containing SiO2 sodium selenite, and citric acid may support the healing of conspicuous cytological findings and clearance of hr-HPV.

Hystoroscopy Assisted Non-Suture Cold-Knife Conization and Cold-Knife Conization in Patients with CINIII: A Comparative Study

167 patients of cervical conization were performed for CINIII in a 34-month study. Conization include Cold-Knife Conization (CKC) (n=120) and hystoroscopy assisted non-suture cold-knife conization (HCKC) (n=47). Both groups were similar in terms of age, indications for conization, and size of cervical cone specimens. Comparing with CKC group, HCKC group was done in a significantly shorter time (28.52±9.80 minutes vs. 52.19±18.09 minutes) (P<0.05), with less intraoperative bleeding of HCKC group (44.89±10.81 ml vs. 51.50±19.94 ml) (P<0.05), shorter hospitalization time (6.89±2.11 days vs. 7.63±1.40 days) (P<0.05, and higher HPV clearance rate (100% vs. 87.13%) (P<0.05) than CKC group. While, the difference in the cure rate (95% vs. 91%) (P>0.05) , the recurrence rate (0% vs. 1%) (P>0.05) and the postoperative complication (P>0.05) between HCKC group and CKC group were not significant. The margins of the excision were checked by rapid histologic examination, showing the incidence of residual CIN was lower in HCKC group (8.51%) than in CKC group (9.17%) (P>0.05). Our study indicates that HCKC is a better treatment than CKC in the management of patients with CINIII.

Factors associated with persistence and clearance of high-risk oral HPV among participants in the HPV Infection in Men (HIM) study

Background HPV attributable oropharyngeal cancer (HPV-OPC) incidence is increasing in many high-income countries among men. Factors associated with oral HPV persistence, the precursor of HPV-OPC, is unknown. Data from the HPV Infection in Men (HIM) study which followed participants &gt;7 years were utilized to examine rates of persistence and associated factors. Methods Oral gargle samples from 3095 HIM study participants were HPV genotyped using SPF10 PCR-DEIA-LiPA25 assay. Oral HPV persistence for individual and grouped high-risk HPV types among 184 men positive for any high-risk HPV at their oral baseline visit was assessed at 6-month intervals. Factors associated with grouped high-risk HPV and HPV16 persistence were examined using logistic regression. Kaplan-Meier curves were constructed to examine median time to HPV clearance overall, and by selected risk factors. Results Among the 7 HPV vaccine types, HPV33 had the longest median duration (7.6 months) followed by HPV16 and HPV45 (6.4 months). 10-30% of oral high-risk HPV infections persisted &gt;24 months. Six months persistence of oral high-risk HPV infections was positively associated with age and gingivitis and negatively associated with lifetime number of sexual partners, while twelve months persistence was only inversely associated with lifetime number of sexual partners. Oral HPV16 persistence was positively associated with baseline HPV16 L1 antibody status. Conclusions 18% of HPV16 infections persisted beyond 24 months potentially conferring higher risk of HPV-OPC among these men. Older age appears to be an important and consistently reported factor associated with oral high-risk HPV persistence. More studies among healthy men are required to understand the progression of oral HPV infection to HPV-OPC.

p16/ki67 and E6/E7 mRNA Accuracy and Prognostic Value in Triaging HPV DNA-Positive Women

Background The study presents cross-sectional accuracy of E6 and E7 (E6/E7) mRNA detection and p16/ki67 dual staining, alone or in combination with cytology and human papillomavirus (HPV)16/18 genotyping, as a triage test in HPV DNA-positive women and their impact on cervical intraepithelial neoplasia (CIN2+) overdiagnosis. Methods Women aged 25-64 years were recruited. HPV DNA-positive women were triaged with cytology and tested for E6/E7 mRNA and p16/ki67. Cytology positive women were referred to colposcopy, and negatives were randomly assigned to immediate colposcopy or to 1-year HPV retesting. Lesions found within 24 months since recruitment were included. All P values were 2-sided. Results 40 509 women were recruited, and 3147 (7.8%) tested HPV DNA positive; 174 CIN2+ were found: sensitivity was 61.0% (95% confidence interval [CI] = 53.6 to 68.0), 94.4% (95% CI = 89.1 to 97.3), and 75.2% (95% CI = 68.1 to 81.6) for cytology, E6/E7 mRNA, and p16/ki67, respectively. Immediate referral was 25.6%, 66.8%, and 28.3%, respectively. Overall referral was 65.3%, 78.3%, and 63.3%, respectively. Cytology or p16/ki67, when combined with HPV16/18 typing, reached higher sensitivity with a small impact on referral. Among the 2306 HPV DNA-positive and cytology-negative women, relative CIN2+ detection in those randomly assigned at 1-year retesting vs immediate colposcopy suggests a -28% CIN2+ regression (95% CI = -57% to +20%); regression was higher in E6/E7 mRNA-negatives (Pinteraction = .29). HPV clearance at 1 year in E6/E7 mRNA and in p16/ki67 negative women was about 2 times higher than in positive women (Pinteraction &lt; .001 for both). Conclusions p16/ki67 showed good performance as a triage test. E6/E7 mRNA showed the highest sensitivity, at the price of too high a positivity rate to be efficient for triage. However, when negative, it showed a good prognostic value for clearance and CIN2+ regression.