DOI: 10.5061/dryad.cvdncjth8
title: Data from: Vaginal bacteria-derived extracellular vesicles diffuse through human cervicovaginal mucus to enable microbe-host signaling

description: The composition of the vaginal microenvironment has significant
 implications for gynecologic and obstetric outcomes. Where a
 Lactobacillus-dominated microenvironment is considered optimal, a
 polymicrobial environment is associated with increased risk for female
 reproductive diseases. Recent work examined bacteria-derived extracellular
 vesicles (bEVs) as an important mode of microbe-host communication that
 may influence reproductive outcomes. However, in order to communicate with
 female reproductive tissues, bEVs must penetrate the protective
 cervicovaginal mucus barrier. We demonstrate increased diffusion of bEVs
 compared to whole bacteria. Additionally, we evaluate the uptake of bEVs
 by, and the resulting effects on, human vaginal epithelial, endometrial,
 and placental cells, highlighting potential mechanisms of action by which
 vaginal dysbiosis contributes to gynecologic and obstetric diseases. Taken
 together, our work demonstrates the ability of bEVs to mediate female
 reproductive outcomes and highlights their potential as therapeutic
 modalities for treating dysbiosis and dysbiosis-associated diseases in the
 female reproductive tract.
description: # **bEV Diffusion and Microbe-Host Response** --- [Access this dataset on
 Dryad](doi.org/10.5061/dryad.cvdncjth8) We have submitted our raw data,
 including bacterial extracellular vesicle (bEV) size distribution
 histograms (**bEV_diffusion_host_response_bEV_Histogram_data_clean.csv**),
 additional characterization
 (**bEV_diffusion_host_response_bEV_Characterization_data_clean.csv**),
 cervicovaginal mucus (CVM) sample information
 (**bEV_diffusion_host_response_CVM_SampleInfo.csv**), 1-second mean
 squared displacements (MSD) per particle
 (**bEV_diffusion_host_response_CVM_1sMSD.csv**), cell viability
 (**bEV_diffusion_host_response_bEV_viability_data_clean.csv**), particle
 uptake (**bEV_diffusion_host_response_bEV_uptake_data_clean.csv**), and
 cytokine response
 (**bEV_diffusion_host_response_bEV_Cytokine_data_clean.csv**). Removed
 outliers can be found in the following associated CSV files: *
 **bEV_diffusion_host_response_bEV_Histogram_removed_outliers.csv** *
 **bEV_diffusion_host_response_bEV_Characterization_removed_outliers.csv**
 * **bEV_diffusion_host_response_bEV_uptake_removed_outliers.csv** *
 **bEV_diffusion_host_response_bEV_viability_removed_outliers.csv** *
 **bEV_diffusion_host_response_bEV_Cytokine_removed_outliers.csv** --- ##
 **Descriptions** ### **General Acronyms** * *LC*: Lactobacillus crispatus
 * *LI*: Lactobacillus iners * *GV*: Gardnerella vaginalis * *MM*:
 Mobiluncus mulieris * *WB*: whole bacteria * *bEV*: bacterial
 extracellular vesicle --- ### **Cell Line Information** * *VK2/E6E7*:
 vaginal epithelium, ATCC CRL-2616 * *Ishikawa*: endometrium, Millipore
 Sigma 99040201 * *BeWo-b30*: placental trophoblast, ATCC CCL-98 --- ###
 **Value Denotation** * *GRUBB*: Value removed as an outlier based on
 Grubbs’ test * *NA*: No applicable data * In removed_outlier.csv files,
 *NA* may indicate that the original value was not an outlier and can be
 found in the corresponding `data_clean.csv` file --- ###
 **bEV_diffusion_host_response_bEV_Histogram_data_clean.csv** * Histogram
 of the distribution of bEV size * First column: size in nm * Consecutive
 columns: independent replicates * First row: bEV species * Second row:
 replicate number --- ###
 **bEV_diffusion_host_response_bEV_Histogram_removed_outliers.csv** * Same
 formatting as the clean file * Lists replicates removed due to Grubbs’
 outlier test --- ###
 **bEV_diffusion_host_response_bEV_Characterization_data_clean.csv** *
 Compiled bEV characteristics by species * First column: bEV species *
 Second column: measurement type and units * Particle concentration
 (bEV/mL), normalized to initial culture volume * Size (nm) * ZP (mV), zeta
 potential * Protein content (µg/1e8 bEVs) * Consecutive columns:
 replicates R1, R2, …, R6 --- ###
 **bEV_diffusion_host_response_bEV_Characterization_removed_outliers.csv**
 * Same formatting as the clean file * Lists replicates removed via Grubbs’
 test --- ### **bEV_diffusion_host_response_CVM_SampleInfo.csv** * Lists
 the average 1-second MSD for all particle types (bEV and WB) * Lists pH
 per CVM sample * First column: sample number (as in publication) * First
 row: species name and WB/bEV classification * Last column: pH per sample
 --- ### **bEV_diffusion_host_response_CVM_1sMSD.csv** * Lists the 1-second
 MSD for each individual particle tracked * First row: sample number *
 Second row: species name * Third row: bEV vs WB * Subsequent rows:
 1-second MSD for every analyzed particle --- ###
 **bEV_diffusion_host_response_bEV_uptake_data_clean.csv** * Percentage of
 internalized bEVs after a dosage of 5000 bEV/cell * First column: cell
 line * Second column: bEV species * Third column: timepoints (h) *
 Consecutive columns: replicates --- ###
 **bEV_diffusion_host_response_bEV_uptake_removed_outliers.csv** * Same
 formatting as the clean file * Lists replicates removed via Grubbs’ test
 --- ### **bEV_diffusion_host_response_bEV_viability_data_clean.csv** *
 First column: cell line * Second column: bEV species * Third column:
 treatment concentration * Consecutive columns: replicates --- ###
 **bEV_diffusion_host_response_bEV_viability_removed_outliers.csv** * Same
 formatting as the clean file * Lists replicates removed via Grubbs’ test
 --- ### **bEV_diffusion_host_response_bEV_Cytokine_data_clean.csv** *
 First column: cell line * Second column: cytokine measured * Third column:
 treatment * Consecutive columns: replicates --- ###
 **bEV_diffusion_host_response_bEV_Cytokine_removed_outliers.csv** * Same
 formatting as the clean file * Lists replicates removed via Grubbs’ test