dendritic cell generation medium  (PromoCell)

Journal: Frontiers in Immunology

Article Title: Mucosal and systemic immune responses after a single intranasal dose of nanoparticle and spore-based subunit vaccines in mice with pre-existing lung mycobacterial immunity

doi: 10.3389/fimmu.2023.1306449

Figure Lengend Snippet: Poly(I:C) improves the adjuvanticity of inactivated spores and YC-NaMA in human monocyte-derived dendritic cells. (A) CD14+ monocytes were magnetically enriched from healthy donor PBMCs After 6 days of differentiation, moDCs were either left unstimulated, or treated with LPS, poly(I:C) or vaccine delivery systems with or without poly (I:C). After 48 hours, cells were surface stained for activation induced markers (MHC-I, MHC-II, CD80, CD86, CD40 and CCR7) for FACS analysis. Culture supernatants were also obtained to quantify secreted inflammatory cytokines (IL-1β, IFN-α2, IFNγ, TNFα, IL-6, IL-18, IL-10, IL-17A, IL-12p70, IL-23, IL-33, CCL2 and CXCL8). (B) Heatmap showing the log10 fold increase in cell-surface and secreted soluble mediators for each treatment group relative to unstimulated group. (C) Heatmap showing P values comparing YC-Nama or Spore with and without Poly(I:C). Data is obtained from the average of two healthy donors. Two-way ANOVA followed by Tukey’s multiple comparisons test was used to compare the groups. Asterisks depicted in A are significant differences from unstimulated. Asterisks in B show significant difference between two groups. P<0.05 = *; P<0.01 = **; P<0.0001 = ****.

Article Snippet: To differentiate CD14+ monocytes to immature moDCs, we used a commercially available dendritic cell generation medium (PromoCell ® , C-28050).

Techniques: Derivative Assay, Staining, Activation Assay