Journal: NPJ Vaccines
Article Title: Systemic prime mucosal boost significantly increases protective efficacy of bivalent RSV influenza viral vectored vaccine
doi: 10.1038/s41541-024-00912-1
Figure Lengend Snippet: a Cytokine responses in CD4 + and CD8 + splenocytes that were separately stimulated with influenza A (H1N1) NP + M1- and RSVF-spanning peptides; % cytokine + T cells were determined through intracellular staining. Basal frequencies of cytokine + T cells (unstimulated sample) were subtracted from stimulated sample frequencies. Median responses in each group are displayed as the top lines of bars on graphs on the left-hand side of the figure, with symbols representing individual mice, grey-shaded bars prime-boost regimens and white bars prime-only regimens. b Cytokine responses in CD4 + and CD8 + lung cells harvested from mice, separately stimulated with influenza A (H1N1) NP + M1- and RSVF-spanning peptides. c Vaccination schematic for the continued assessment of the cellular immunogenicity of ChAdOx1-NP + M1-RSVF. d IFNγ responses in splenocytes stimulated with NP + M1- and RSVF-spanning peptides (IFNγ spot-forming cells (SFCs)/million splenocytes), as measured by IFNγ enzyme-linked immunosorbent spot (ELISpot) assay. e Total counts of CD8 + T RM cells (left), as well as relative counts of CD8 + T EM and T RM cells (right) in lungs post-vaccination (all non-antigen-specific). T RM cells were defined as CD8 + CD44 + CD62L - CD103 + CD69 + , and negative for intravenous (IV) circulatory CD3 + T cell stain. T EM cells were defined as CD8 + CD44 + CD62L - CD127 + , and negative for IV circulatory CD3 + T cell stain. Group differences of data in ( a ), ( b ), ( d ) and ( e ) were analysed using non-parametric Kruskal-Wallis tests (*= p < 0.05, **= p < 0.01, ***= p < 0.001). For all boxplots, whisker endings represent upper and lower extremes, the box bounds represent upper and lower quartiles, respectively, and the central line represents the group median.
Article Snippet: PVDF-membrane ELISpot plates (Millipore) were coated with 5μg/mL anti-mouse IFNy (Cat: 3321-2 A, AN18 (Mabtech)), 50 μL per well.
Techniques: Staining, Immunopeptidomics, ELISpot Assay, Enzyme-linked Immunospot, Whisker Assay
Millipore is a verified supplier 
