Journal: bioRxiv
Article Title: Tissue resident cells differentiate S. aureus from S. epidermidis via IL-1ß following barrier disruption in healthy human skin
doi: 10.1101/2024.02.19.580932
Figure Lengend Snippet: ( A ) Intracellular CFU counts of S. epidermidis 1457 (filled circles) and S. aureus USA300 (empty circles) 3 h p.i. of tape-stripped corneocytes, human keratinocytes (HEKa) and fibroblasts (HDFn). Bars represent means ± SD, dots show individual data points (corneocytes and fibroblasts n=3; keratinocytes n=5; 3 technical replicates). Dotted line = initial inoculum. Significance was determined by a two-way ANOVA with Tukey’s correction for multiple comparisons. Significance is denoted by **** p ≤ 0.0001. ( B ) IL-8 response displayed as x-fold change over uninfected controls measured by ELISA in cell culture supernatants of S. epidermidis 1457 or S. aureus USA300 infected corneocytes, keratinocytes or fibroblasts. Dot plot shows individual data points (corneocytes and fibroblasts n = 3; keratinocytes n = 5; 3 technical replicates each). Significance was determined by a two-way ANOVA with Tukey’s correction for multiple comparisons. Significance is denoted by *** p ≤ 0.001 or **** p ≤ 0.0001. ( C ) Microscopic detection of intracellular S. aureus GFP in cell monolayers of tape-stripped corneocytes, human keratinocytes (HEKa) or fibroblasts (HDFn); uninfected CTR vs 3h infection. Immunostaining of f-actin (red) and nuclei (blue) in HEKa and HDFn, as well as autofluorescence staining of corneocytes (red). Scale bars = 50µm.
Article Snippet: The human dermal fibroblasts cell line (HDFn) (Gibco, Thermo Fischer Scientific Cat# C0045C) was cultured in Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F-12) (Gibco, Thermo Fischer Scientific Cat# 31330038) supplemented with L-glutamine (Thermo Fischer Scientific Cat# A2916801), 15 mM HEPES and 5% charcoal-stripped fetal bovine serum (FBS) (Gibco, Thermo Fischer Scientific Cat# 12676029) at 37°C with 5% CO2.
Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Infection, Immunostaining, Staining