Fig. 4: Magnitude and polyfunctionality of cellular responses elicited by SMARRT.RSV.preF in NHPs.

a RSV.F specific IFNγ secreting cells in PBMCs of immunized animals (n = 4 biologically independent animals per group) represented as number of Spot Forming Units (SFU) per million cells. The dotted line represents the positivity threshold of the assay which is set at 50 SFU and the red line indicates the geometric mean response. Background subtracted response of each animal (gray circles) overtime is shown. Comparisons were made between baseline response to weeks 8 and 12. b CD4 (square) and CD8 (circle) memory T-cells gated on Live CD45⁺CD28^±D95⁺ positive for CD107a or IFNγ or TNFα or IL2 stimulated with RSV.F peptide pool were identified using intracellular cytokine staining. After subtracting the corresponding response in the medium stimulated sample of each animal, values are shown with a threshold represented by the dotted line and median response by the red line. Due to insufficient number of cells, data from an RSV pre-exposed animal immunized with 10 mcg SMARRT.RSV.preF group is not available (b–d). Polyfunctional subsets of CD4 and CD8 T-cells by Boolean gating and after background subtraction were subsequently analyzed using SPICE at week 4 post-immunization. Bar plots represent background-subtracted median frequency of cells in each subset for each animal (circles). Pie chart wedges represent the functional subsets producing different combinations of cytokines i.e., 4+, 3+, 2+ and monofunctional subsets indicated by the color coding under “Pie”, while the surrounding pie arcs represent total median level of each analyte. Statistical analysis was done on square root transformed values. b–d Comparisons between SMARRT.RSV.preF immunized groups was determined with ANOVA (TOBIT model) and adjusted for multiple comparisons with Bonferroni correction. Source data are provided as a Source Data file.