vk2 e6e7 human vaginal epithelial cell line  (ATCC)

Journal: bioRxiv

Article Title: Drug screen reveals new potent host-targeted antivirals against Mpox virus

doi: 10.1101/2025.05.02.651913

Figure Lengend Snippet: (A) Representative immunofluorescence images of MPXV-infected VK2/E6E7 cells treated with seven selected drug compounds at varying concentrations (1.657, 0.828, 0.414, 0.207, 0.103, and 0.051 µM) for 48 hours post-infection (hpi). Blue and red indicate nuclei and MPXV-A4L antigen, respectively. Scale bar = 100 µm. (B) The dose-response curves of the indicated compounds are shown. The percentage of MPXV-Dose–response curves showing percent inhibition of MPXV infection in VK2/E6E7 cells treated with the indicated compounds. Cells were infected with MPXV and fixed at 48 hours post-infection (hpi) for immunofluorescence analysis. Infection levels were calculated as the ratio of MPXV-positive cells (red) to total DAPI-stained nuclei (blue), normalized to vehicle-treated controls. Percent inhibition values were derived as 1 minus the normalized infectivity. Nonlinear regression was used to calculate IC 50 values, which are displayed along with curve fit R values. The dotted horizontal line indicates 50% inhibition. Data represent the mean ± standard deviation from triplicate wells.

Article Snippet: The VK2/E6E7 (VK2) human vaginal epithelial cell line (ATCC, CRL-2616) was maintained in keratinocyte serum-free medium (KSFM) supplemented with 0.1 ng/mL human recombinant epidermal growth factor (EGF), 0.05 mg/mL bovine pituitary extract (BPE), and 100 U/mL penicillin-streptomycin.

Techniques: Immunofluorescence, Infection, Inhibition, Staining, Derivative Assay, Standard Deviation

Journal: Microorganisms

Article Title: Bacillus coagulans LMG S-24828 Impairs Candida Virulence and Protects Vaginal Epithelial Cells against Candida Infection In Vitro

doi: 10.3390/microorganisms12081634

Figure Lengend Snippet: Evaluation of C. albicans (Ca) adhesion capacity to a vaginal epithelial cell monolayer in the presence of B. coagulans (Bc) ( A ). The histogram graph shows the average % ± SEM of fungal adhesion inhibition exerted by B. coagulans (Bc). Data are from three independent experiments. ( B ) Assessment of B. coagulans (Bc) capacity to co-aggregate with C. albicans (Ca) after 1 h of co-incubation. Boxes in the heatmap represent the score assigned to each sample in three independent experiments—0: no aggregation; 1: aggregates with small clusters; 2: aggregates with larger numbers of yeasts; 3: clumps visible with the naked eye containing large numbers of yeast cells; 4: maximum score for large clumps visible with the naked eye in the well center. ( C – E ) Effect of B. coagulans (Bc) on C. albicans (Ca) hyphal formation upon 4 h of co-incubation. Hyphal fragments were optically counted by fluorescent microscopy imaging. The fungal cell wall was stained with Uvitex 2B fluorescent dye. ( C ) The bars chart reports the mean percentage ± SEM of hyphal fragments counted in three different fields from three independent experiments. Statistical analysis was performed by the unpaired, two-tailed Student t -test. Ca vs. Bc + Ca * p < 0.05. ( D ) The heatmap shows the % of hyphal fragments counted in each field; the squares’ color indicates the abundance of hyphae in the field (red: high % hyphal fragments; green: low % hyphal fragments). ( E ) Representative images from fluorescence microscopy analysis are shown from C. albicans (Ca) or C. albicans plus B. coagulans (Bc + Ca) taken at 40× magnification.

Article Snippet: Two different human epithelial cell lines were employed: the A-431 cell line from vaginal epithelial squamous cell carcinoma (ATCC CLR-1555) and the CaCo-2 cell line from colon–rectal adenocarcinoma (ATCC HTB-37).

Techniques: Inhibition, Incubation, Microscopy, Imaging, Staining, Two Tailed Test, Fluorescence

Journal: Microorganisms

Article Title: Bacillus coagulans LMG S-24828 Impairs Candida Virulence and Protects Vaginal Epithelial Cells against Candida Infection In Vitro

doi: 10.3390/microorganisms12081634

Figure Lengend Snippet: ( A ) Percentage of vaginal cell damage pre-colonized or not by B. coagulans (Bc) for 6 h and infected for further 18 h with C. albicans (Ca). The chart reports the average percentage of cell damage ± SEM of triplicate samples from three different experiments. Statistical analysis was performed by the unpaired, two-tailed Student t -test. Ca vs. Bc + Ca * p < 0.05. ( B ) Production of β-defensin-2 by vaginal epithelial cells pre-colonized or not by B. coagulans (Bc) for 6 h and infected for further 18 h with C. albicans (Ca). Uninfected cells (Ctrl) and cells colonized by the bacterium without C. albicans were also included in the experiments. The graph reports the mean ± SEM from three independent experiments. Statistical analysis was performed by the one-way ANOVA test followed by the uncorrected Fisher’s LSD test. Untreated cells vs. Bc pre-colonized cells * p < 0.05.

Article Snippet: Two different human epithelial cell lines were employed: the A-431 cell line from vaginal epithelial squamous cell carcinoma (ATCC CLR-1555) and the CaCo-2 cell line from colon–rectal adenocarcinoma (ATCC HTB-37).

Techniques: Infection, Two Tailed Test

Journal: Microorganisms

Article Title: Bacillus coagulans LMG S-24828 Impairs Candida Virulence and Protects Vaginal Epithelial Cells against Candida Infection In Vitro

doi: 10.3390/microorganisms12081634

Figure Lengend Snippet: Evaluation of antifungal effect permanency upon B. coagulans removal. C. albicans (Ca) ( A ) and C. parapsilosis (Cp) ( B ) metabolic activity quantification after being incubated with B. coagulans or sterile medium for 24 h and subsequent fungal isolation and cultivation for 24 h in the lack of bacteria. The graphs show the mean OD at 492 nm wavelength ± SEM from triplicate sample of three different experiments. Statistical analysis was performed by the one-way ANOVA test followed by the uncorrected Fisher’s LSD test. ns = not significant. ( C ) Capacity of B. coagulans spores to germinate on intestinal epithelial cells CaCo-2. Bacterial spores were seeded on an intestinal epithelial cell monolayer of CaCo-2 and incubated at 37 °C + 5% CO 2 for 24 h. After incubation, monolayers were photographed (upper images) and subsequently lysed. A Gram staining was then performed to visualize the presence of germinated B. coagulans (Bc) (lower images).

Article Snippet: Two different human epithelial cell lines were employed: the A-431 cell line from vaginal epithelial squamous cell carcinoma (ATCC CLR-1555) and the CaCo-2 cell line from colon–rectal adenocarcinoma (ATCC HTB-37).

Techniques: Activity Assay, Incubation, Sterility, Isolation, Bacteria, Staining

Journal: Cell Reports

Article Title: Vaginal epithelial dysfunction is mediated by the microbiome, metabolome, and mTOR signaling

doi: 10.1016/j.celrep.2023.112474

Figure Lengend Snippet:

Article Snippet: VK2/E6E7 immortalized human epithelial vaginal cell line , ATCC , Cat# CRL-2616; RRID: CVCL_6471.

Techniques: Virus, Bacteria, Recombinant, Software

Journal: mBio

Article Title: A New Phenotype in Candida -Epithelial Cell Interaction Distinguishes Colonization- versus Vulvovaginal Candidiasis-Associated Strains

doi: 10.1128/mbio.00107-23

Figure Lengend Snippet: C. albicans hyphal fragment production. Data in the graphs show the mean percentage ± standard deviation (SD) of hyphal fragments produced by VVC and colonizing C. albicans strains after 24 h of culture in RPMI 1640 (protocol i) (A), in RPMI or RPMI plus 10% FCS in the presence or absence of Lactobacillus rhamnosus (L.r.) (protocol ii) (B), or during infection of a monolayer of A431 cells (protocol iii) (C). Statistical analysis was performed according to Student's t test (A and C) and Kruskal-Wallis test, followed by Dunn’s multiple-comparison test or one-way ANOVA, followed by Tukey’s multiple-comparison test (left and right graphs of panel B, respectively). Panels D and E show representative images of hyphal segments from the 01887 (D) and 14314 (E) strains from experimental protocol ii, grown in sgRPMI. Arrowheads indicate the septa separating one hyphal segment from another. The data come from at least 3 biological replicates.

Article Snippet: The human A431 vaginal epithelial cell line, obtained from ATCC, was grown in Dulbecco’s modified Eagle’s medium (DMEM) plus 10% defined fetal bovine serum (FBS) (HyClone, Logan, UT, USA), gentamicin (50 mg/mL) (Bio Whittaker, Verviers, Belgium), ciprofloxacin (Ciproxin) (2 mg/mL) (ICN), and l -glutamine (2 mM) (EuroClone, Milan, Italy).

Techniques: Standard Deviation, Produced, Infection, Comparison

Journal: mBio

Article Title: A New Phenotype in Candida -Epithelial Cell Interaction Distinguishes Colonization- versus Vulvovaginal Candidiasis-Associated Strains

doi: 10.1128/mbio.00107-23

Figure Lengend Snippet: CA01887 and CA14314 differentially activate type I interferon, integrin, and ferroptosis pathways. A431 vaginal epithelial cells were challenged with either CA01887 or CA14314 for 24 h. Differential RNA-seq was performed on two biological replicates of mock-infected versus infected challenges and analyzed with DEseq2 and Ingenuity Pathway Analysis. (A to C) Differentially regulated genes from three pathways with divergent responses upon C. albicans challenge are shown with statistically significant upregulation (yellow), significant downregulation (blue), or no significant change (white). The range for each is shown below (values are the log 2 ratio of fungal challenge to mock challenge). The cutoff for significant changes was an adjusted P value ( P adj ) of <0.01. (D to F) Summary of IFNAR inhibition of shedding for several VVC and colonizing strains. Shown is the mean percentage of change ± SD in CFU from at least 3 different experiments after 24 h of infection of the vaginal epithelial monolayer with VVC or colonizing strains in the presence or absence of neutralizing IFNAR antibody (Ab). All experiments used the shedding protocol. (D) C. albicans CFU shed in the supernatants (cells in suspension); (E) C. albicans CFU attached to vaginal epithelium (exfoliated, adherent, and loosely adherent cells); (F) overall C. albicans CFU for each infection (all cells). Red dots indicate the strains CA01887 and CA14314, which were used in the original RNA-seq experiments. The statistical comparisons between VVC and colonizing strains were performed according to the unpaired Student's t test. P values of <0.05 (*) were considered significant. n.s., not significant ( P > 0.05).

Article Snippet: The human A431 vaginal epithelial cell line, obtained from ATCC, was grown in Dulbecco’s modified Eagle’s medium (DMEM) plus 10% defined fetal bovine serum (FBS) (HyClone, Logan, UT, USA), gentamicin (50 mg/mL) (Bio Whittaker, Verviers, Belgium), ciprofloxacin (Ciproxin) (2 mg/mL) (ICN), and l -glutamine (2 mM) (EuroClone, Milan, Italy).

Techniques: RNA Sequencing, Infection, Inhibition, Suspension