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. 2022 Mar 31;52(6):936–945. doi: 10.1002/eji.202249823

Figure 2.

Figure 2

Intranasal immunization with ChAdOx1‐S induces a superior mucosal immunity in the lungs compared to intramuscular immunization. (A) BALB/c mice were vaccinated via intranasal (IN) or intramuscular (IM) route with ChAdOx1‐S. (B) At 6 weeks after the first immunization, mice were boosted, and at 9 weeks mucosal, Spike‐specific immune response in BALs was assessed by ELISA (n = 3‐4). Dotted lines indicate the median absorbance value of unimmunized mice sera. Neutralization titers were determined by in vitro neutralization assay (all groups n = 5). Dotted lines indicate the median value of IU/mL of unimmunized mice sera. Dots represent individual data points. (C) Eight weeks after the first immunization, spleen and lung homogenates were restimulated with Spike‐specific peptide KNKCVNFNF (S535‐543). The responding CD8 T cells were identified by intracellular staining for accumulated IFN‐γ. Bars represent group means overlaid with individual data points (n = 10). (D) Tissue‐resident phenotypes of antigen‐experienced CD8 T cells (CD8+CD44+) were assessed by staining for CD69 and/or CD103. Data were analyzed by one‐way ANOVA followed by Tukey's multiple comparison test; p values indicate significant differences (*p < 0.05; **p < 0.01, ***p < 0.001, ****p < 0.0001). All experiments (B–D) have been repeated two to three times. IU: international units.