Figure 5

Oral treatments with LL 301 restore salivary flow in diseased SjS mice. (A) Groups of 16 week-old SjS females (8/group) were orally treated with PBS, 5 × 10⁹ CFUs WT LL, or 5 × 10⁹ CFUs LL 301, and a second dose administered 4 weeks later. SFR measurements were taken (B) prior and (C) after second treatment; ***P < 0.001, *P < 0.05 versus the indicated group. Study was terminated 24 weeks of age, and SMGs were formalin fixed and stained with hematoxylin and eosin to determine extent of inflammatory cell infiltration. Images of stained tissues at × 20 magnification were examined for infiltrated regions using the Aperio ImageScope software as described in Fig. 3. Focus score of infiltrates were determined by using (D) the number of foci and (E) focus size in area; *P < 0.05 versus WT LL-treated mice are shown. One-way ANOVA followed by Dunnett’s multiple comparisons test was performed. Splenic lymphocytes were analyzed for percentages of (F) Foxp3⁺ CD4⁺ and (G) Foxp3⁺ CD25⁺ CD4⁺ T cells, and for the percentages of (H) IL-17⁺ and (I) IFN-γ⁺ CD4⁺ T cells; *P < 0.05 relative to PBS- or WT LL-dosed mice are shown. (J,M) Purified HNLN, (K,N) MLN, and (L,O) splenic lymphocytes from PBS-, WT LL-, and LL 301-treated mice were stimulated with anti-CD3 and anti-CD28 mAbs for 4 days. Culture supernatants were analyzed for production of (J–L) IL-17 and (M–O) IFN-γ by cytokine-specific ELISAs. Depicted are the means ± SEM; ****P < 0.0001, **P < 0.01, *P < 0.05 versus the indicated group.