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. Author manuscript; available in PMC: 2023 May 10.
Published in final edited form as: Immunity. 2022 Apr 27;55(5):895–911.e10. doi: 10.1016/j.immuni.2022.04.001

Figure 7. LasB-deficient P. aeruginosa have reduced adjuvant activity to stimulate eosinophil recruitment and mucin production.

Figure 7.

(A and B) Cell frequency (A) and numbers (B) of eosinophils (CD45+MHCII–CD11b+CD64-Siglec-F+) and neutrophils (CD45+MHCII–CD11b+CD64-Ly6G+) out of CD45+ cells in the BAL of mice immunized with OVA +/− WT or ΔLasB PA01 (combined of 3 experiments yielding 9–11 mice per group).

(C and D) H&E (C) and PAS (D) staining in the lungs of mice immunized as in (A and B) (representative of 3 experiments, 4 mice per group, scale bar = 500 μm).

(E) Relative mRNA expression of type 2 cytokines (IL-4, 5, 13), eotaxins (Ccl11, 24) and Muc5AC in the lungs of mice immunized as in (A and B) (combined of 2 experiments yielding 7–9 mice per group).

(F) ELISA of IFN-γ amounts in the BAL of mice immunized with OVA +/− WT or ΔLasB PA01 (combined of 2 experiments yielding 5–7 mice per group).

(G) (Top) Representative fast protein liquid chromatography (FPLC) profile (n=3) of Muc5AC before and after incubation with PA01 and (bottom) growth curves of PA01 on Muc5AC (growth curve data are mean ± SEM, n=3; FPLC data are the mean of n=3 traces).

(H) (Top) PA01 growth +/− LasB (3 μg/ml) and/or phosphoramidon (50 mM) and (bottom) WT and ΔLasR PA01 growth +/− LasB (3 μg/ml) on human sputum samples for 12 hr at 37 °C (data are mean ± SEM, n=4).

mRNA expression was measured relative to Rpl13a. Data were analyzed by one-way ANOVA with Tukey’s multiple comparisons test. *p < 0.05; **p < 0.01; ***p < 0.001. See also Figures S6 and S7.