Fig. 7: In vivo antigen delivery recalls local and systemic memory responses.
Six months after the final immunization, non-human primates were given an in vivo antigen recall in the skin and lungs with H107, or a control protein, as shown in (Fig. 6). A Serum H107-specific IgG (left) and IgA (right) antibody levels as measured by ELISA before (pre) and post in vivo antigen recall of H107/CAF®-immunized Non-human primates. Symbols represent mean ± SD of arbitrary units, n = 5/grp. B, C PBMCs from control and H107/CAF®-immunized NHPs (n = 5/grp) were assessed by ELISpot assay pre- and post in vivo antigen recall. PBMCs were stimulated ex vivo for H107-specific (B) IFNγ and (C) IL-17A production as in Fig. 5. D The percentage of CD4 T cells isolated from skin biopsies collected from control antigen (Ctrl Ag) and H107 protein ID injection sites that express IFNγ, TNF, IL-2, IL-17, and/or IL-13 after ex vivo stimulation with H107 protein, as assessed by intracellular cytokine staining (ICS), n = 5/grp. E Sample contour plots of ICS analysis of CD4 T cells isolated from the BAL of a representative H107/CAF®10b-immunized animal 8 days after in vivo antigen (left), and the resultant summary data of the total % H107-specific cytokine-expressing BAL CD4 T cells. Bars represent median ± IQR, min. and max. Symbols, individual animals, n = 5/grp, n = 4 CAF®09lo. P values, two-sided Mann-Whitney U tests. Kruskal-Wallis test p = 0.0097. F The percentage of cytokine-producing CD4 T cells from PBMC samples collected pre- and post-in vivo antigen recall assessed IFNγ, TNF, IL-2, IL-17, and/or IL-13 after ex vivo stimulation with H107 protein. B–D, F Symbols and lines, individual animals n = 5/grp. P values within groups from two-sided paired T test. G Principle component analysis (PCA) of H107-specific CD4 T cells from BAL (red) and PBMC (blue) for IL-17 and combinatorial IFNγ/TNF/IL-2 expression. PC1 and PC2 loading axes shown. Symbol, individual H107/CAF®-immunized animals, n = 5/grp. Source data are provided as a Source Data file.
