Journal: Microorganisms

Article Title: Modulation of C. albicans-Induced Immune Response in Vaginal Epithelial Cells by Garcinoic Acid

doi: 10.3390/microorganisms12122455

Figure Lengend Snippet: Figure 5. Proposed model of mechanistic aspects of GA immuno-modulatory effects in vaginal epithelial cells exposed to C. albicans infection. The MKK3/6-p38/SAPK pathway and its regulatory interplay with pro-survival MAPK ERK1/2 are molecular targets of GA, with modulating activity on IL-6, IL-1 α, and IL-1β levels secreted by the infected vaginal epithelium. The effect of GA on these cytokines may preserve the immune-metabolic properties of the vaginal epithelium and support post-infection recovery and the restoration of cellular homeostasis. (A) Mechanism of action of C. albicans on vaginal epithelial cells. (B) Effect of pre-treatment with GA on vaginal cells infected with C. albicans. The dotted line in the arrows indicates that there is a trend toward a non-significant reduction in the proposed event; the solid line indicates a significant reduction/inhibition; red arrows indicate increased production; blue arrows indicate decreased secretion. The image was produced by using BioRender software (https://www.biorender.com; license number: C81D6A59-0002).

Article Snippet: The A-431 cell line from vaginal epithelial squamous cell carcinoma (ATCC CLR-1555) was cultured in Dulbecco’s Modified Eagle Medium (DMEM; Invitrogen, Carlsbad, CA, USA) supplemented with 1% penicillin/streptomycin solution (Euroclone, Milan, Italy) and 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Infection, Activity Assay, Inhibition, Produced, Software

Journal: Nature microbiology

Article Title: Candida pathogens induce protective mitochondria-associated type I interferon signalling and a damage-driven response in vaginal epithelial cells.

doi: 10.1038/s41564-021-00875-2

Figure Lengend Snippet: Fig. 1 | Pathogenicity patterns of four Candida species in the in vitro vaginal epithelial infection model. a, Adhesion, determined as the percentage of Candida cells from the original inoculum that adhered to vaginal epithelial cells at 1 h post-infection; invasion, determined as the percentage of Candida cells that invaded the vaginal epithelial cells at 3 h post-infection; hyphal length (µm) recorded at 3 h post-infection; necrotic damage, measured by the quantification of LDH activity in the supernatant and presented as a percentage respective to total lysis (maximum damage control) at 24 h post-infection. All values are presented as mean ± s.d. of n = 3 independent in vitro infection experiments. b, Generation times of Candida species in YPD or RPMI 1640 medium (used for vaginal epithelial cell infections) measured in 24-h growth curves. All values are presented as the mean ± s.d. of n = 5 independent experiments. No statistically significant difference in growth between species in neither YPD nor RPMI was observed (one-way ANOVA with Greenhouse– Geisser correction and Tukey’s multiple comparisons test). c, Micrographs of Candida morphology at 3 h post-infection and confluent biofilms at 24 h post-infection on vaginal epithelial cells. Micrographs are representative of n = 3 independent experiments with similar results.

Article Snippet: Primary human vaginal epithelial cells (catalogue no. PCS480-010) were obtained from ATCC and cultured in vaginal epithelial cell basal medium (catalogue no. PCS-480-030; ATCC), supplemented with components from the Vaginal Epithelial Cell Growth Kit (catalogue no. PCS-480-040; ATCC).

Techniques: In Vitro, Infection, Activity Assay, Lysis, Control

Journal: Nature microbiology

Article Title: Candida pathogens induce protective mitochondria-associated type I interferon signalling and a damage-driven response in vaginal epithelial cells.

doi: 10.1038/s41564-021-00875-2

Figure Lengend Snippet: Fig. 6 | Type-I IFN signalling increases epithelial resistance and suppresses innate immune activation. a, Epithelial damage caused by C. albicans 24 h post-infection after RNA interference for selected ISGs (MX2, CMPK2, IFI6). Transfection with control siRNA, consisting of a scrambled sequence that will not lead to the specific degradation of any mRNA, was used as a control (dotted line). b, Epithelial damage caused by C. albicans 24 h post-infection without and with 0.1 ng ml−1 of IFN-β and the addition of anti-IFNAR antibody. c, Levels of IL-6, IL-1β and IL-1α secretion by neutrophils incubated with supernatants from infected epithelial cells and by epithelial cells infected with Candida species 24 h post-infection. d, Levels of IL-8 secretion by infected epithelial cells 24 h post-infection with or without the addition of anti-IFNAR antibody. e, Levels of IL-8 secretion by neutrophils after 24-h incubation with supernatants from epithelial cell infections with or without the addition of an anti-IFNAR antibody. Control supernatants of Candida cells alone were included to ensure that the neutrophils responded to secretions of epithelial cells rather than the fungus (blue bars). f, Schematic model of the proposed role of the type I IFN pathway in immune regulation and protection against Candida infection. All values are presented as the mean ± s.d. of n = 3 independent experiments (except n = 4 for IL-8 production by epithelial cells on C. albicans infection). Statistical significance is indicated as *P ≤ 0.05, **P ≤ 0.01, ****I ≤ 0.0001 (one-way ANOVA with Dunnett’s multiple comparisons test (a) or Tukey’s multiple comparisons test (b,d)). Credit: graphics in f adapted from Servier under a Creative Commons licence CC BY 3.0.

Article Snippet: Primary human vaginal epithelial cells (catalogue no. PCS480-010) were obtained from ATCC and cultured in vaginal epithelial cell basal medium (catalogue no. PCS-480-030; ATCC), supplemented with components from the Vaginal Epithelial Cell Growth Kit (catalogue no. PCS-480-040; ATCC).

Techniques: Activation Assay, Infection, Transfection, Control, Sequencing, Incubation

Journal: Polymers

Article Title: The Effect of β-Glycerophosphate Crosslinking on Chitosan Cytotoxicity and Properties of Hydrogels for Vaginal Application

doi: 10.3390/polym7111510

Figure Lengend Snippet: Figure 9. Results of the cytotoxicity studies (% of cell viability, apoptosis and necrosis referred to non-treated cells) using flow cytometry (A–C) or fluorescent microscopy assay (D–F) after 4 h of exposure of human vaginal epithelial cells CRL 2616 with controls (as described in Table 5) (A,D), two concentrations (0.1; 1.0 mg¨ mL´1) of unmodified or β-GP crosslinked LMw (B,E) or MMw CS (C,F) (mean ˘ S.D.; n = 3).

Article Snippet: Normal human vaginal epithelial cells (CRL 2616; VK2/E6E7) were purchased from American Type Culture Collection (Manassas, VA, USA).

Techniques: Cytometry, Microscopy

Journal: Polymers

Article Title: The Effect of β-Glycerophosphate Crosslinking on Chitosan Cytotoxicity and Properties of Hydrogels for Vaginal Application

doi: 10.3390/polym7111510

Figure Lengend Snippet: Figure 10. Representative fluorescence microscopy images of viable (A); apoptotic (B) and necrotic (C) human vaginal epithelial cell CRL 2616 incubated with CS/lGP at a concentration of 1.0 mg¨ mL´1

Article Snippet: Normal human vaginal epithelial cells (CRL 2616; VK2/E6E7) were purchased from American Type Culture Collection (Manassas, VA, USA).

Techniques: Microscopy, Incubation, Concentration Assay