Fig. 3: Local immune response in nasal washes and NALTs in BALB/c mice immunized with rNA-N1-MPP vaccine adjuvanted with BDX100 or BDX301 adjuvants. | npj Vaccines

Fig. 3: Local immune response in nasal washes and NALTs in BALB/c mice immunized with rNA-N1-MPP vaccine adjuvanted with BDX100 or BDX301 adjuvants.

From: Immunogenicity and protective efficacy of an intranasal neuraminidase-based influenza vaccine with bacterial cell membrane-derived adjuvants

Fig. 3

a Representative flow cytometry plots showing the percentage of GCB and Tfh populations in NALTs (gated on live CD3‒CD19+ and CD3+CD19‒CD4+ cells, respectively). b Percentage of GCB and Tfh populations in NALTs 10 days after the booster immunization (n = 5). c Level of IgA and IgG in the nasal washes obtained from the immunized animals 10 days after the booster immunization. Dots represent individual AUC values; bars represent average values (n = 5). Groups were compared using ANOVA followed by the Tukey posthoc test. P-values for the groups with statistically significant differences are shown on the plots. d Differential expression of phenotype markers in NALT subpopulations identified with PhenoGraph clustering algorithm. T‑cell clusters (1–9, T) and B‑cell clusters (10–20) can be segregated based on CD3/CD4/CD8 and CD19 expression. Cluster 1: Activated effector memory CD4 T cells (CD4⁺CD44+CD62L‒CD69+); Cluster 2: Tfh (CD4⁺PD‑1+CD185+); Cluster 3: CD4 TRM (CD4⁺CD103+CD69+); Cluster 4: Naïve CD4 (CD4⁺CD62L+CD44‒); Cluster 5: Central memory CD4 T-cells (CD4⁺CD62L+CD44+); Cluster 6: Effector/effector‑memory (E/EM) CD4 T-cells (CD4⁺CD62L‒CD44lo); Cluster 7: Effector‑memory CD8 T-cells (CD8⁺CD62L‒CD44+); Cluster 8: Central‑memory CD8 T-cells (CD8⁺CD62L+CD44+); Cluster 9: CD8⁺CD103+CD69‒; Cluster 10: Isotype-switched (IS) memory B cells (CD19⁺CD273+CD80+IgDlo); Cluster 11: Unswitched/activated memory B cells (CD19⁺CD273‒CD80+IgDlo); Cluster 12: GCB (CD19+CD38‒GL7+); Cluster 13: Early memory (Early mem., CD19⁺CD273‒CD80+IgD+IgM+); Cluster 14: Naïve B-cells (CD19⁺CD62L‒IgD+IgM+); Cluster 15: Naïve CD62L+ B-cells (CD19⁺CD62L+IgD+IgM+); Cluster 16: Transitional plasmablast-like (PB-like, CD19⁺CD138+CD103+); Cluster 17: PD1+ B-cells; Cluster 18: Isotype‑switched (IS) B-cells (CD19⁺CD273‒CD80‒IgD‒IgM‒); Cluster 19: Early GCB (CD19+CD38+GL7+); Cluster 20: Activated B-cells (CD19⁺CD69‒IgD+IgM+). e PhenoGraph cluster percentage from the total number of live cells in NALTs 10 days after the booster immunization. Samples were combined for percentage calculation. f UMAP projections showing the distribution of PhenoGraph clusters in NALTs of mice 10 days after the booster immunization with adjuvanted or nonadjuvanted N1-MPP or PBS. g UMAP projections showing the differential expression of phenotype markers on NALT cells. h Percentage of PhenoGraph clusters for which the statistical differences were demonstrated between the experimental groups in the ANOVA test (n = 5). Groups were compared using ANOVA followed by the Tukey posthoc test. P-values for the groups with statistically significant differences are shown on the plots.

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