Viral infection in presence of each CVL or control buffer.

*<p>IQR interquartile range.</p

Proteins identified in more than one active, but not in the inactive CVL samples by LC-ESI-MS/MS.

<p>Proteomic analysis was performed on the starting material (A), flow through (B), and retentate (C) after fractionation with a Centricon 50 kDa spin column from an active CVL pool (1) or two different individual CVL (2, 3). Sample 4 was separated on a gel filtration column and the starting material (A) and the fraction with the greatest bactericidal activity (D) were evaluated.</p

Endogenous anti-HIV activity in CVL collected in the absence or following coitus.

<p>TZM-bl cells were infected with 10³ TCID₅₀ HIV-1_BaL in the presence of Visit 1 (baseline) or Visit 2 (postcoital) CVL or control buffer (saline with 1.5 mg/ml bovine serum albumin). Mock infected cells were included as controls. Results are mean RLU obtained from 2 independent experiments, each conducted in triplicate.</p

Demographic and clinical data from 10 couples.

<p>F, female; M, male; min, minutes.</p>*<p>Time (minutes) between intercourse and CVL collection for Visit 2 and time after gel application of CVL collection for Visits 3 and 4.</p

Loss in the anti-HSV activity in CVL collected following PRO 2000 gel application in the absence or following coitus.

<p>CaSki cells were infected with serial 10-fold dilutions of HSV-2(G) mixed with each CVL sample. After incubation for 2 h, the inoculum was removed by washing and the cells were overlaid with fresh media. Infection was monitored by counting plaques after 48 h. Results are viral titer (pfu/ml) calculated from 2 independent experiments where each sample was tested in duplicate. The upper panel shows results for each subject for Visit 1 (baseline) versus Visit 4 (post-gel), middle panel for Visit 4 (post-gel) versus Visit 3 (post-gel and postcoital) and lower panel for Visit 2 (postcoital) versus Visit 3 (post-gel and postcoital).</p

Lactobacillus Proteins Are Associated with the Bactericidal Activity against <em>E. coli</em> of Female Genital Tract Secretions

<div><h3>Background</h3><p>Female genital tract secretions are bactericidal for <em>Escherichia (E.) coli ex vivo</em>. However, the intersubject variability and molecules that contribute to this activity have not been defined.</p> <h3>Methods</h3><p>The bactericidal activity and concentration of immune mediators in cervicovaginal lavage (CVL) collected from 99 healthy women were determined.</p> <h3>Results</h3><p>CVL reduced the number of <em>E. coli</em> colonies by 68% [−26, 100] (median [range]). CVL were active against laboratory and clinical isolates of <em>E. coli,</em> but were inactive against Lactobacillus species. Bactericidal activity correlated with the concentration of protein recovered (p<0.001), but not with cytokines, chemokines or antimicrobial peptides. Four CVL with>90% inhibitory activity (active) and two with<30% activity were subjected to MS/MS proteomic analysis. 215 proteins were identified and six were found exclusively in active samples. Four of these corresponded to <em>Lactobacillus crispatu</em>s or <em>jensenii</em> proteins. Moreover, culture supernatants from <em>Lactobacillus jensenii</em> were bactericidal for <em>E. coli.</em></p> <h3>Conclusion</h3><p>Both host and commensal microbiota proteins contribute to mucosal defense. Identification of these proteins will facilitate the development of strategies to maintain a healthy vaginal microbiome and prevent colonization with pathogenic bacteria such as <em>E. coli</em> that increase the risk for urinary tract infections, preterm labor and perinatal infection.</p> </div

Distribution of CVL <i>E. coli</i> bactericidal activity from a cohort of 99 healthy females.

<p>Each CVL or control fluid (200 µg/mL of bovine serum albumin in normal saline) was mixed with ∼10⁹ cfu of <i>E. coli</i> for 2 h. The mixture was then diluted to yield 800–1000 cfu on control plates and plated in duplicate on tryptic soy agar plates. Colonies were counted after 24 h incubation and the mean percentage inhibition relative to the control plates was determined.</p

Outcome variables including bactericidal activity, vaginal wall pH, and concentration of immune mediators and bivariate regression estimate for <i>E. coli</i> inhibition.

*<p>β coefficients and p values for immune mediators refer to log transformed values.</p>#<p>Modeled as dichotomized variable: Interferon α (>or≤12.25) and Interferon γ (>or≤0.05).</p

MS/MS data of the S-layer protein of <i>L. crispatus</i>.

<p>MS/MS of the precursor doubly charged ion at <i>m/z</i> 608.05 matching to the S-layer protein peptide DAILQLGGDALK from <i>L. crispatus</i> (A). The other matching peptides are highlighted in yellow and the green highlights are residues that have undergone deamidation (B).</p

Distribution of demographic, sexual behaviors and history of STI and bivariate regression estimate for <i>E. coli</i> bactericidal activity.

#<p>modeled as dichotomized variable ≥1 partner(s) vs. no partners.</p