Fig. 3: The STING signalling pathway is required for the adenovirus vector-induced adjuvant effect.

a, Endpoint titres of anti-RBD IgG in serum samples from WT or transgenic (Tlr9^−/−, Tlr2^−/−, Myd88^−/−, Nlrp3^−/−, Casp1^−/−, Il1b^−/−, Sting^−/−, Cd8a^−/−) mice immunized with Ad5_BA.5 alone or Ad5_BA.5 plus RBD_BA.5-HR (n = 6 mice per group). b, Endpoint titres of anti-RBD IgA in BALF samples from WT or Sting^−/−mice immunized with Ad5_BA.5 alone or Ad5_BA.5 plus RBD_BA.5-HR (n = 6 mice per group). c, Endpoint titres of sera anti-RBD IgG from WT or transgenic mice immunized with Ad5_Empty plus RBD_BA.5-HR. WT mice intranasally vaccinated with naked RBD_BA.5-HR were used as control (n = 6 mice per group). d, Endpoint titres of BALF IgA from WT or Sting^−/−mice immunized with Ad5_Empty plus RBD_BA.5-HR (n = 6 mice per group). e, Absolute number of antigen-experienced CD8⁺ T_RM cells in BALF from vaccinated WT, Myd88^−/− or Sting^−/− mice (n = 6 mice per group). f, t-SNE maps were generated by concentrating CD3⁺ T cells in the BALF from WT or Sting^−/− mice vaccinated with Ad5_BA.5 plus RBD_BA.5-HR, and heat map projections of CD69 and CD103 expression on t-SNE maps are shown (n = 3 mice each group). Analysis was carried out using FlowJo v.10. g,h, Percentage of spike-specific CD8⁺ (g) and CD4⁺ (h) memory T cells in lung tissue of vaccinated WT, Myd88^−/− or Sting^−/− mice (n = 6 mice per group). i, Percentage of antigen-specific IFNγ-producing CD8⁺ T cells in lung from mice vaccinated with Ad5_Empty + RBD_BA.5-HR (n = 6 mice per group). Data are presented as geometric mean ± s.d. (a–d), and as mean with s.e.m. (e,g–i). P values were determined using two-tailed unpaired Student’s t-tests (a,b), one-way ANOVA followed by Dunnett’s multiple comparisons test (c) or Tukey’s multiple comparisons test (d), two-way ANOVA followed by Sidak’s multiple comparisons test (e–h), and one-way ANOVA followed by Tukey’s multiple comparisons test (i).

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