human a431 vaginal epithelial cell line  (ATCC)

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