Evidence for cGAS-STING signaling in the female genital tract resistance to Chlamydia trachomatis infection

Sexually transmitted Chlamydia trachomatis can ascend to the upper genital tract due to its resistance to innate immunity in the lower genital tract. C. trachomatis can activate cGAS-STING signaling pathway in cultured cells via either cGAS or STING. The current study was designed to evaluate the role of the cGAS-STING pathway in innate immunity against C. trachomatis in the mouse genital tract. Following intravaginal inoculation, C. trachomatis significantly declined by day 5 following a peak infection on day 3 while the mouse-adapted C. muridarum continued to rise for >1 week, indicating that C. trachomatis is susceptible to the innate immunity in the female mouse genital tract. This conclusion was supported by the observation of a similar shedding course in mice deficient in adaptive immunity. Thus, C. trachomatis can be used to evaluate innate immunity in the female genital tract. It was found that mice deficient in either cGAS or STING significantly increased the yields of live C. trachomatis on day 5, indicating an essential role of the cGAS-STING signaling pathway in innate immunity of the mouse genital tract. Comparison of live C. trachomatis recovered from different genital tissues revealed that the cGAS-STING-dependent immunity against C. trachomatis was restricted to the mouse lower genital tract regardless of whether C. trachomatis was inoculated intravaginally or transcervically. Thus, we have demonstrated an essential role of the cGAS-STING signaling pathway in innate immunity against chlamydial infection, laying a foundation for further illuminating the mechanisms of the innate immunity in the female lower genital tract.

SCREENING OF INFERTILE FEMALES FOR LOWER GENITAL TRACT INFECTIONS

Introduction: Infertility has been known to cause serious social and emotional problems worldwide. Besides other causes of female infertility, the role of female reproductive tract infection is well recognized. Lower genital tract infection, be it symptomatic or asymptomatic, need to be diagnosed and treated properly. In view of this our study was done. Aim & Objectives: To evaluate the bacteriological profile of lower genital tract in infertile females. Methodology: It was a cross sectional type of study. After taking consent, three swabs (high vaginal swab, endocervical swab and swab from lateral vaginal wall) were taken from 100 infertile women. A questionnaire covering demographic data, menstrual history, medical history, history of infertility, etc. was completed for each of the participants. Isolation and identification of the isolates were done as per conventional techniques. Antibiotic Susceptibility Testing was done for the aerobic isolates as per CLSI guidelines. Results: In our study, 47% of females were asymptomatic and majority them showed positive microbiological growth. E.coli and S.aureus were the most common aerobic isolates and Prevotella spp. was the most common anaerobic isolate. Majority of the anaerobes were associated with bacterial vaginosis. Majority of our isolates were susceptible to Gentamicin. Conclusion: The absence of clinical symptoms does not rule out the possibility of an ongoing acute inflammatory state due infective agents. Hence, both asymptomatic and symptomatic females should be screened for lower genital tract infections as the consequences may lead to infertility.

Case Report: Intra-Tumoral Vaccinations of Quadrivalent HPV-L1 Peptide Vaccine With Topical TLR-7 Agonist Following Recurrence: Complete Resolution of HPV-HR-Associated Gynecologic Squamous Cell Carcinomas in Two Patients

The human papilloma virus (HPV) high-risk variants (HPV-HR) such as HPV16 and HPV18 are responsible for most HPV related cancers, including anogenital and head and neck cancers. Here, we present two patients with HPV-HR-associated gynecological malignancies who, after failing radiation therapy, were treated with experimental salvage immunotherapy regimen resulting in complete, durable responses in both patients. Each patient was diagnosed with recurrent, radiation-refractory, HPV-HR positive, squamous cell carcinoma of the lower genital tract. Patient A was a 90-year-old, African American, with metastatic vulvar cancer to the right inguinal-femoral triangle and pulmonary metastases. Patient B was a 41-year-old, Caucasian, with a central-recurrence of cervix cancer. Each patient received at least two intratumoral quadrivalent HPV-L1 vaccine (Gardasil™) injections and daily topical TLR-7 agonist (imiquimod) to the tumor surface 2 weeks apart. This combination of intratumoral vaccinations and topical TLR-7 agonist produced unexpected complete resolution of disease in both patients. The importance of radiation therapy, despite being considered a treatment failure by current definitions, cannot be understated. Radiation therapy appears to have offered a therapeutic immune advantage by modifying the tumor microenvironment. This immune protocol has potential to help patients with advanced HPV-HR-related malignancies previously considered incurable.

Predictive Risk Factors for Mild and Severe Germinal Matrix Hemorrhage and Associated Neurodevelopmental Prognosis: A Retrospective Study

Aims: To compare commonly mentioned risk factors between mild germinal matrix hemorrhage-Intraventricular hemorrhage (GMH-IVH) (grade I & II) and severe GMH-IVH (grade III & IV) and to study the long-term neurodevelopmental outcomes in relation to severe GMH-IVH. Study Design: Retrospective cohort study. Place and Duration of Study: Neonatal intensive care unit, King Fahad University Hospital, between 2000 and 2020. Methodology: We included 54 premature infants at ≤36 weeks of gestation and with birth weight <2500g admitted to our neonatal intensive care unit. Premature neonates were divided into two subgroups: mild GMH-IVH (grade I and II) and severe (grade III and IV). We investigated the risk factors and neurodevelopmental outcomes in association with GMH-IVH. Results: Amnionitis (OR: 1.190, 95% CI 0.515-2.749), lower genital tract infection (OR: 1.190, 95% CI 0.515-2.749), antenatal infection (OR: 1.406, 95% CI 0.866-2.283), gestational diabetes mellitus (OR: 1.815, 95% CI 1.410-2.337), usage of inotropes (OR: 1.731, 95% CI 1.348-2.222), APGAR score <7 (OR: 0.806, 95% CI 0.493-1.316), birth trauma (OR: 1.767, 95% CI 1.396-2.236), catecholamines (OR: 1.470, 95% CI 0.903-2.393), intubation (OR: 1.300, 95% CI 0.686-2.464), asphyxia (OR: 1.135, 95% CI 0.718-1.794), Abnormal coagulation (OR: 1.197, 95% CI 0.756-1.896), congenital heart disease (OR: 1.727, 95% CI 1.124-2.653), low hematocrit (OR: 1.140, 95% CI 0.688-1.889), resuscitation (OR: 1,193, 95% CI 0.748- 1.904) and ventriculoperitoneal (VP) shunt as a prognosis of hydrocephalus (P-value: 0.005) all showed a higher incidence with severe GMH-IVH Conclusion: Amnionitis, lower genital tract infection, antenatal infections, GDM, usage of inotropes, APGAR score <7, birth trauma, catecholamines, intubation, asphyxia, resuscitation, abnormal coagulation parameters, congenital heart disease, low hematocrit and hydrocephalus with VP shunt were higher in severe GMH-IVH.

An Insight into Vaginal Microbiome Techniques

Background: There is a unique microbial community in the female lower genital tract known as the vaginal microbiota, which varies in composition and density and provides significant benefits during pregnancy, reproductive cyclicity, healthy newborn delivery, protection from preterm birth, infections such as UTIs, bacterial vaginosis, and so on, and improves the efficacy of treatments for vaginal cancers. Methods: It is necessary to know how the vaginal microbiome is composed in order to make an accurate diagnosis of the diseases listed above. A microbiome’s members are difficult to classify, and the way microbial communities function and influence host–pathogen interactions are difficult to understand. More and more metagenomic studies are able to unravel such complexities due to advances in high-throughput sequencing and bioinformatics. When it comes to vaginal microbiota research, we’ll be looking at the use of modern techniques and strategies that can be used to investigate variations in vaginal microbiota in order to detect diseases earlier, better treat vaginal disorders, and boost women’s health. Discussion: The discussed techniques and strategies may improve the treatment of vaginal disorders and may be beneficial for women’s overall health.

Cervicovaginal microbiota predicts Neisseria gonorrhoeae clinical presentation

Neisseria gonorrhoeae infection of the female lower genital tract can present with a spectrum of phenotypes ranging from asymptomatic carriage to symptomatic cervical inflammation, or cervicitis. The factors that contribute to the development of asymptomatic or symptomatic infections are largely uncharacterized. We conducted a pilot study to assess differences in the cervicovaginal microbial community of patients presenting with symptomatic vs. asymptomatic N. gonorrhoeae infections to a sexually transmitted infections (STI) clinic. DNA was isolated from cervicovaginal swab specimens from women who tested positive for N. gonorrhoeae infection using a clinical diagnostic nucleic acid amplification test. We performed deep sequencing of 16S ribosomal RNA gene amplicons, followed by microbiome analyses with QIIME, and species-specific real-time PCR to assess the composition of microbial communities cohabitating the lower genital tract with the infecting N. gonorrhoeae. Specimens collected from asymptomatic individuals with N. gonorrhoeae infection and no co-infection with Chlamydia trachomatis and/or Trichomonas vaginalis carried Lactobacillus-dominant microbial communities more frequently than symptomatic patients without co-infection. When compared to asymptomatic individuals, symptomatic women had microbial communities characterized by more diverse and heterogenous bacterial taxa, typically associated with bacterial vaginosis (BV) (Prevotella, Sneathia, Mycoplasma hominis and Bacterial Vaginosis-Associated Bacterium-1 (BVAB1)/”Candidatus Lachnocurva vaginae). Both symptomatic and asymptomatic N. gonorrhoeae patients with additional STI co-infection displayed a BV-like microbial community. We used a murine model of N. gonorrhoeae infection in mice pre-colonized with Lactobacillus crispatus to test whether pre-existing L. crispatus was protective from N. gonorrhoeae colonization or whether N. gonorrhoeae infection could drive the loss of L. crispatus during infection. Vaginal infection with either N. gonorrhoeae strain 1291 or an isogenic mutant known to exhibit lower inflammatory had no impact on Lactobacillus burden recovered from the mice. These data taken together suggest that Lactobacillus-dominant vaginal microbial community may protect individuals from developing symptoms during lower genital tract infection with N. gonorrhoeae.

132 Early Establishment of the Human Gut Microbiome—a Tale of Moms, Their Diets, and Babes Guts

Human microbial communities are characterized by their metagenomic and metabolic diversity, which varies by distinct body sites and influences human physiology. We are only beginning to characterize the complex set of interactions which alters both community membership and function in early development. With respect to the potential source of microbiota at birth, it has been generally assumed that the majority of seeding microbes originate from the maternal lower genital tract, with microbiota ascending into the otherwise sterile intrauterine. However, we and subsequently others have recently demonstrated that (1) the vaginal and gut microbiome communities are distinctly structured in pregnancy, and (2) the intrauterine environment and the fetus is in fact not sterile, but rather harbors a low-abundance microbiome which varies by several measured exposures, and (3) the maternal diet during both gestation and lactation, and notably a high fat diet, has a particularly strong impact on the developing and early in life microbial community structure. We have taken two dynamic approaches to answering these questions in our studies. First, we use large and robust longitudinal cohorts of maternal-infant dyads collected across gestation and into infancy to gain deeper insight into both source and sink of the early developmental microbiome and its role on determining length of gestation. Second, we utilize our well established primate models of maternal high fat dietary exposure, both in the absence and presence of maternal obesity, to determine the impact of maternal diet on both the microbiome and the resultant offspring metabolic phenotype.