Journal: Scientific Reports
Article Title: Inflammatory and antimicrobial properties differ between vaginal Lactobacillus isolates from South African women with non-optimal versus optimal microbiota
Figure Lengend Snippet: Inflammatory cytokine production by VK2 cells in response to different vaginal Lactobacillus species. Lactobacillus isolates, including L. crispatus (n = 11), L. jensenii (n = 14), L. johnsonii (n = 5), L. mucosae (n = 15), L. plantarum (n = 2), L. ruminis (n = 5), L. salivarius (n = 2) and L. vaginalis (n = 10), were adjusted to 4.18 × 10 6 colony forming units (CFU)/ml in antibiotic free keratinocyte serum free media before being incubated with VK2 cells for 24 hours at 37 °C with 5% CO 2 . Cytokine concentrations were measured in the culture supernatants using Luminex. ( A ) Stacked bars showing median concentrations of each cytokine interleukin (IL)-1α, IL-1β, IL-6, IL-8, IFN-γ-inducible protein (IP)-10, macrophage inflammatory protein (MIP)-1α, MIP-1β, MIP-3α and regulatory IL-1 receptor antagonist (RA). ( B ) Stacked bars showing median concentrations of each cytokine excluding IL-1RA (IL-1α, IL-1β, IL-6, IL-8, IP-10, MIP-1α, MIP-1β and MIP-3α).
Article Snippet: Vaginal epithelial cell stimulation and measurement of cytokine concentrations Vaginal epithelial cells (VK2/E6E7 ATCC CRL-2616), that closely resemble the tissue of origin, were maintained in complete keratinocyte serum free media (KSFM) supplemented with 0.4 mM calcium chloride, 0.05 mg/ml of bovine pituitary extract, 0.1 ng/ml human recombinant epithelial growth factor and 50 U/ml penicillin and 50 U/ml streptomycin (Sigma-Aldrich, St. Louis, Missouri) as described previously.
Techniques: Incubation, Luminex