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. 2023 Jun 29;8:96. doi: 10.1038/s41541-023-00691-1

Fig. 9. Longevity of cellular immune responses induced by IN immunization.

Fig. 9

The durability of the antigen recall responses induced by immunization was measured by comparing the levels of secreted cytokines from splenocytes (SPL) or cervical lymph node isolates (cLN) from immunized mice in response to ex vivo stimulation with 5 μg RBD for 72 h. SPL and cLN were isolated from young (8-week old) mice given three IN immunizations with 10 μg RBD with either NE or NE/IVT either 2 or 25 weeks after the last immunization (week 10 or week 33, respectively). Levels of secreted cytokines were measured in the cell supernatant by multiplex immunoassay relative to unstimulated cells for (a) IFN-γ, b IL-2, c IP-10, d TNF-α, e IL-4, f IL-5, g IL-13, h IL-6, i IL-17A, and j IL-10. (data shown as mean ± SEM (n = 5/grp)); (*p < 0.05, **p < 0.01 as compared between week 10 and week 33 for each adjuvant group by Mann–Whitney U test).