Extended Data Fig. 2: Lymph node dendritic cells analysis.

a, The quantitative results of the absolute total cell number of LNs (blank (8–10 wo), blank (80–90 wo), Vac (80–90 wo), n = 4; Vac (8–10 wo), n = 5, biological replicates). P values: Vac (80–90 wo) versus blank (80–90 wo) P = 0.4487; Vac (8–10 wo) versus blank (8–10 wo) *P = 0.5483; blank (80–90 wo) versus blank (8–10 wo) P = 0.6497; Vac (80–90 wo) versus Vac (8–10 wo) *P = 0.0489. b, c, The proportion and absolute number of immune cells (CD45⁺) in the LNs (blank (8–10 wo), Vac (80–90 wo), blank (80–90 wo), n = 4; Vac (8–10 wo), n = 5, biological replicates). (b) P values: Vac (80–90 wo) versus blank (80–90 wo) P = 0.1138; Vac (8–10 wo) versus blank (8–10 wo) P = 0.0815; blank (80–90 wo) versus blank (8–10 wo) **P = 0.0029; Vac (80–90 wo) versus Vac (8–10 wo) **P = 0.0028. (c) P values: Vac (80–90 wo) versus blank (80–90 wo) P = 0.6238; Vac (8–10 wo) versus blank (8–10 wo) P = 0.1625; blank (80–90 wo) versus blank (8–10 wo) P = 0.0816; Vac (80–90 wo) versus Vac (8–10 wo) *P = 0.0408. d, Correlations between the absolute number of CCR7⁺ DCs and T_FH cells (n = 22, biological replicates; P < 0.0001). e, Correlations between the number of CCR7⁺ DCs, T_FH cells and antibody titers (n = 22, biological replicates). The number of CCR7⁺ DCs significantly correlated with both antibody titers (R² = 0.7785, P < 0.0001) and T_FH cell numbers (R² = 0.7876, P < 0.0001). Similarly, T_FH cell numbers showed a strong positive correlation with antibody titers (R² = 0.7490, P < 0.0001). f, LNs immune cell clustering genes. g, h, t-SNE map for regrouping analysis of CD45⁺ cells and corresponding quantitative results. i, j, t-SNE map for regrouping analysis of DCs and corresponding quantitative results. k, Highlight t-SNE map of representative genes in DCs. l, Violin plot of genes identified for DC subset. m, n, The absolute number of cDC2 (CD11b⁺XCR1⁻MHCII⁺CD11c⁺Epcam⁻CD45⁺) and correlations between cDC2 and serum anti-RBD IgG binding titers in the LNs (n = 4, biological replicates per group). *P = 0.0456. o, p, The absolute number of cDC1 (XCR1⁺CD11b⁻MHCII⁺CD11c⁺Epcam⁻CD45⁺) and correlations between cDC1 and antibody titers (Vac (8–10 wo), n = 5; Vac (80–90 wo), n = 4, biological replicates). q, r, The absolute number of LCs (CD207⁺Epcam⁺MHCII⁺CD11c⁺CD45⁺) and correlations between LCs and antibody titers (Vac (8–10 wo), n = 5; Vac (80–90 wo), n = 4, biological replicates). s, t, The absolute number mig cDC1 (XCR1⁺CD11b⁻MHCII^highCD11c⁺Epcam⁻CD45⁺) and correlations between mig cDC1 and antibody titers (n = 4, biological replicates per group). u, v, The absolute number res cDC1 (XCR1⁺CD11b⁻MHCII^intCD11c⁺Epcam⁻CD45⁺) and correlations between res cDC1 and antibody titers (Vac (8–10 wo), n = 5; Vac (80–90 wo), n = 4, biological replicates). w, x, The absolute number res cDC2 (CD11b⁺XCR1⁻MHCII^intCD11c⁺Epcam⁻CD45⁺) and correlations between mig cDC1 and antibody titers (Vac (8–10 wo), n = 5; Vac (80–90 wo), n = 4, biological replicates). Data are means ± SEM. Statistical analysis was performed using Student’s t-test (two-tailed). NS, not significant.

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