Karen Smith-McCune

The contraceptive effectiveness of intrauterine devices (IUDs) has been attributed in part to a foreign body reaction in the endometrium. We performed this study to better understand mechanisms of action of contraceptives of by studying their effects on endometrial and cervical transcriptomes. We collected endometrial and cervical biopsies from women using the levonorgestrel-releasing intrauterine system (LNG-IUS, n = 11), copper intrauterine device (cu-IUD, n = 13) or levonorgestrel-containing combined oral contraceptives (COC, n = 12), and from women not using contraceptives (control group, n = 11). Transcriptional profiling was performed with Affymetrix arrays, Principal Component Analysis and the bioconductor package limma. In endometrial samples from cu-IUD users, there were no genes with statistically significant differential expression compared to controls. In LNG-IUS users, 2509 genes were differentially expressed and mapped predominantly onto immune and inflammatory pathways. The cervical samples showed no statistically significant differential gene expression compared to controls. Hormonal and copper IUDs have significantly different effects on the endometrial transcriptome, with the LNG-IUS transcriptome showing pronounced inflammation and immune activation compared to controls whereas the cu-IUD transcriptome was indistinguishable from luteal phase endometrium. These findings argue against a foreign body reaction as a common mechanism of action of IUDs.

Progestins are widely used for the treatment of gynecologic disorders and alone, or combined with an estrogen, are used as contraceptives. While their potencies, efficacies and side effects vary due to differences in structures, doses and routes of administration, little is known about their effects on the endometrial transcriptome in the presence or absence of estrogen. Herein, we assessed the transcriptome and pathways induced by progesterone (P₄) and the three most commonly used synthetic progestins, medroxyprogesterone acetate (MPA), levonorgestrel (LNG), and norethindrone acetate (NETA), on human endometrial stromal fibroblasts (eSF), key players in endometrial physiology and reproductive success. While there were similar transcriptional responses, each progestin induced unique genes and biofunctions, consistent with their structural similarities to progesterone (P₄ and MPA) or testosterone (LNG and NETA), involving cellular proliferation, migration and invasion. Addition of estradiol (E₂) to each progestin influenced the number of differentially expressed genes and biofunctions in P₄ and MPA, while LNG and NETA signatures were more independent of E₂. Together, these data suggest different mechanisms of action for different progestins, with progestin-specific altered signatures when combined with E₂. Further investigation is warranted for a personalized approach in different gynecologic disorders, for contraception, and minimizing side effects associated with their use.

Globally, HIV/AIDS is a leading cause of morbidity worldwide among reproductive-aged cisgender women, highlighting the importance of understanding effects of contraceptives on HIV-1 risk. Some observational studies suggest there may be an increased risk of HIV-1 acquisition among women using the long-acting injectable progestin contraceptive, depo-medroxyprogesterone acetate. The potential mechanism of this susceptibility is unclear. There are few data on the role of the upper female reproductive tract in HIV-1 transmission, and the mechanisms of HIV-1 infection are likely to differ in the upper compared to the lower reproductive tract due to differences in tissue composition and variable effects of sex steroids on mucosal immune cell distribution and activity. In this study, we measured the susceptibility of mucosal immune cells from the upper female reproductive tract to HIV-1 entry using the virion-based HIV-1 fusion assay in samples from healthy female volunteers. We studied 37 infectious molecular clones for their ability to fuse to cells from endometrial biopsies in three participants and found that subtype (B or C) and origin of the virus (transmitted founder or chronic control) had little influence on HIV-1 fusion susceptibility. We studied the effect of contraceptives on HIV-1 susceptibility of immune cells from the cervix, endometrium and peripheral blood by comparing fusion susceptibility in four groups: users of the copper intrauterine device (IUD), levonorgestrel-containing oral contraceptive, levonorgestrel-containing IUD and unexposed controls (n = 58 participants). None of the contraceptives was associated with higher rates of HIV-1 entry into female reproductive tract cells compared to control samples from the mid-luteal phase.

Objective To understand glycosylation of endocervical proteins at different times throughout the menstrual cycle in naturally cycling women and in women using hormonal or non-hormonal contraceptive methods, in order to characterize biochemical fingerprints of favorable and unfavorable cervical mucus. Design Lectin/antibody-probed protein blot analysis of endocervical mucus samples collected onto ophthalmologic sponges (wicks) from two groups: a longitudinal cohort of naturally cycling women at three time points in their menstrual cycles (discovery cohort), and a cross-sectional cohort of women on hormonal or non-hormonal contraceptive methods (validation cohort). Setting Participants were recruited from the San Francisco Bay Area from 2010 to 2016. Patient(s) Women with regular cycles not using hormonal or intrauterine device (IUD) contraceptives were recruited for the longitudinal cohort (n = 8). Samples from women using levonorgestrel-containing combined oral contraceptives (n = 16), levonorgestrel containing IUDs (n = 14), copper IUDs (n = 17), depo-medroxyprogesterone acetate (DMPA) (n = 15), and controls (n = 13) were used for validation. Intervention(s) None. Main outcome measure(s) Detection of specific glycosylation patterns on lectin/antibody probed protein blots. Result(s) Two lectins (Lens culinaris agglutinin and Lycopersicon esculentum [tomato lectin]), and the antibody MECA-79 demonstrated consistent cycle-dependent changes in protein binding. The glycan-binding patterns of the levonorgestrel-containing contraceptives were generally similar to each other and to those from women in the luteal phase. The DMPA samples showed slightly different binding patterns. Conclusion(s) We identified molecular signatures of unfavorable mucus from women in the luteal phase and on hormonal contraceptives. Further characterization of these biomarkers may be useful in contraceptive development and in evaluation of infertility.

Problem: There is little information regarding the impact of the intrauterine device on immune parameters of the upper female reproductive tract related to risk of HIV acquisition.Method of Study: We collected cervical and endometrial samples from women using the hormonal intrauterine device to study its effects on endocervical cytokines/chemokine concentrations, phenotypic markers of T cells, responses of endometrial T cells to activation, and alterations of endometrial cellular infiltrates.Results: Hormonal intrauterine device use was associated with: increased concentrations of inflammatory cytokines/chemokines (endocervix); increased coexpression of CXCR4 and CCR5 (endocervix and endometrium); increased coexpression of CD38 and HLADR (endocervix and endometrium); increased intracellular IL‐10 production after T‐cell stimulation (endometrium); and increased density of T cells, most notably regulatory T cells (endometrium).Conclusion: Hormonal intrauterine device use resulted in both inflammatory and immunosuppressive alterations. Further research is needed to determine the significance of these changes for HIV risk.

We assessed the feasibility of a breath test to detect women’s single or concurrent use of vaginal products by adding ester taggants to vaginal gel and condom lubricant. Healthy non-pregnant women were enrolled into a two-day cohort (N = 13) and a single-day cohort (N = 12) in San Francisco. Within each cohort, women were randomized (5:1) to tagged or untagged products, and inserted in a clinical setting: 4 mL of tenofovir placebo gel (ten tagged with 15 mg 2-pentyl acetate; three untagged), and an artificial phallus with a lubricated condom (11 tagged with 15 mg 2-butyl acetate; two untagged), on two separate days (two-day cohort) or concurrently (single-day cohort). Using a portable mini-gas chromatograph, the presence/absence of taggants was determined in breath specimens collected prior to, and at timed intervals following product exposure. Demographic, clinical and product use experience data were collected by structured interview. All participants completed all visits and inserted their assigned products. At 5 min post-insertion, the breath test was 100 % accurate in identifying insertion of the tagged (or untagged) gel and/or condom. The half-life in breath of the two esters tested was N = 4) and scent (N = 5). The tagged products were well tolerated. This breath test has the potential to accurately and objectively monitor adherence to vaginal gel and condom used separately or concurrently. Se evaluó la viabilidad de una prueba de respiración para detectar, solo o concurrente, uso de los productos vaginales por mediante de las mujeres, añadiendo etiquetados éster al gel vaginal y lubricante de condón. Mujeres saludables, no embarazadas, se inscribieron en una cohorte de dos días (N = 13) y una cohorte de un solo día (N = 12) en San Francisco. Dentro de cada cohorte, mujeres fueron aleatorizadas (5:1) a productos, con o sin etiquetas, que fueron insertados dentro de un entorno clínico: 4 ml del gel placebo tenofovir (10 etiquetado con 15 mg de acetato de 2-pentilo; 3 sin etiqueta), y un falo artificial con un condón lubricado (11 etiquetado con 15 mg de 2-acetato de butilo; 2 no etiquetado), en dos días separados (cohorte de dos días), o concurrentemente (cohorte de un solo día). Usando un mini-cromatógrafo de gases portátil, la presencia / ausencia de etiquetados se determinaron en muestras de aliento recogidas antes de y a intervalos de tiempo, después de la exposición del producto. Datos demográficos, clínicos y de la experiencia sobre el uso del producto se recogieron mediante entrevista estructurada. Todos los participantes completaron todas las visitas y insertaron sus productos asignados. A los 5 minutos después de inserción, la prueba de alcoholemia fue 100% preciso en la identificación de la inserción, con o sin etiqueta, del gel y / o condón. El tiempo que tardo para la mitad de los ésters desglosar y ser eliminados del cuerpo por medio del aliento, de los dos ésteres que fueron usados para la prueba, fue <1 hora con una gran variabilidad entre los individuos, indicadores y cohortes. En general, entre los productos relacionados, etiquetados y recibidos, fueron reportados seis eventos adversos leve y dos moderados. Todos fueron transitorios y se resolvieron espontáneamente. Sensaciones adicionales incluyeron sabor en la boca (n = 4) y aroma (n = 5). Los productos etiquetados fueron bien tolerados. Esta respiración de ensayo es capaz de controlar con precisión y objetivamente la adherencia al gel vaginal y condón usado por separado o concurrentemente.