Figure 1.
C57BL/6J and IL-23 deficient mice exhibit comparable survival patterns following IN or ID LVS infection.
Groups of 5 to 10 WT mice and p19M KO mice were infected IN with LVS doses ranging from 101–106 CFU (doses 101 and 106 not shown for clarity). For ID infections, groups of 5 WT mice and p19M KO were infected ID with LVS doses ranging from 104–106. The calculated LD50 for IN and ID infections are shown (inset table). No mice from any ID-infected group succumbed to infection. Results shown are representative of three experiments of similar design and outcome. p19G KO mice were also included in some experiments with similar results (data not shown).
Figure 2.
Bacterial burdens in tissues are comparable between infected WT mice and p19G KO mice infected with LVS ID and IN.
Groups of 3 WT mice or p19G KO mice were infected either ID with 105 LVS (A), or were anesthetized and infected with 2×103 LVS IN (B). Mice were sacrificed at the indicated time points, and lungs, livers, and spleens plated to enumerate CFU. There were no significant differences in organ burdens between WT mice and p19G KO mice for any organ at any time point. Results shown are representative of two experiments of similar design and outcome. p19M KO mice were also included in one experiment with similar results (data not shown).
Figure 3.
Serum antibodies are comparable between WT and p19 KO mice infected with LVS by the intradermal or intranasal route.
Groups of 5 WT or p19M KO mice were infected with 105 LVS ID, or were anesthetized and infected with 2×103 LVS IN. All mice were bled before infection (pre-bleed; pre), and at the indicated time points during and after infection. Sera from individual mice for each group were pooled and assayed for LVS-specific total IgM and IgG antibodies by ELISA. OD is represented on the Y axis, and two-fold serial dilutions are represented on the X axis, where dilution 1 = 1∶40 dilution. A, left panel) IgM was measured in sera from WT mice or p19M KO mice at day 8 after ID infection. A, right panel) IgG was measured from WT mice or p19M KO mice at day 42 after ID infection. B, left panel) IgM was measured in sera from WT mice or p19M KO mice at day 8 after IN infection. B, right panel) IgG was measured from WT mice or p19M KO mice at day 35 after IN infection. On all panels, results from pre-bleeds from WT mice (WT Pre) are also shown; results from KO mouse pre-bleeds are not shown for clarity, but were the same as those for pre-bleeds from WT mice and essentially negative. Results shown are representative of two experiments of similar design and outcome. Similar results were obtained for sera from p19G mice (data not shown).
Table 1.
Similar titers of serum antibodies titers in LVS-vaccinated WT and IL-23 KO mice.
Table 2.
Similar survival of lethal challenge by LVS-vaccinated WT and IL-23 KO mice.
Figure 4.
T cell functions and cytokine production are similar between WT mice and p19G KO mice, regardless of priming route.
WT mice and p19G KO mice were infected with 105 LVS ID or 103 LVS IN. Mice were sacrificed after 8 weeks and splenocytes isolated; splenocytes were also obtained from naïve mice. Bone marrow-derived macrophages from WT mice (shown) or p19G KO mice (not shown) were infected with LVS, and then 5×106 homologous splenocytes were added to triplicate wells of infected macrophages. A) Intracellular bacterial burdens were assessed immediately after infection at 0 hours (not shown) and at 72 hours after infection. B) Supernatants were harvested from cultures at 72 hours after infection, and the levels of RNI measured. Supernatants were also assayed for cytokines by sandwich ELISA, including C) IFN-γ and D) IL-17A. IL-12p40 and IL-12p70 were also measured (data not shown). Results are shown for one representative experiment of three experiments of similar design and outcome. *p value≤0.05 by Student’s t test.