Journal: Nature Communications
Article Title: Augmentation of vaccine-induced humoral and cellular immunity by a physical radiofrequency adjuvant
Figure Lengend Snippet: RF induces low-level local inflammation. Dorsal skin of C57BL/6 mice were exposed to RF or intradermally injected with 20 µl Alum (1:1 volume ratio in PBS), AddaVax (50%, vol/vol in PBS), or MPL (25 µg). Adjuvant-treated and non-treated skins were dissected at indicated times. a , b Heat map of relative cytokine ( a ) and chemokine ( b ) gene expression. Total RNA was extracted followed by reverse transcription and real-time PCR analysis of cytokine and chemokine gene expression using GAPDH as an internal control. The baseline gene expression level was set at 1. c – g Different innate immune cell levels in RF and adjuvant-treated skin. Skin was digested in collagenase D and dispase to prepare single-cell suspensions. Cells were then stained with fluorescence-conjugated antibodies followed by flow cytometry analysis of levels of different cell types: neutrophils ( c ), monocytes ( d ), macrophages ( e ), eosinophils ( f ), and mDCs ( g ) (Supplementary Fig. 2). n = 4. Student’s t -test was used to compare differences between groups at 48 and 96 h. *, p
Article Snippet: Skin was digested in 0.2% collagenase D (Roche Diagnostics, Mannheim, Germany) and 0.6U/ml dispase (Gibco, Invitrogen, Carlsbad, CA) in PBS at 37˚C for 3 h with intermittent vortexing.
Techniques: Mouse Assay, Injection, Expressing, Real-time Polymerase Chain Reaction, Staining, Fluorescence, Flow Cytometry, Cytometry