Figures & data
Figure 7. IgE binding capacity of DN-Dp. N: Native Der p crude extract; DN: DN-Dp dialyzed in PBS buffer. Protein extracts were separated on 12% SDS-PAGE. After electrophoresis, protein extracts were transferred to nitrocellulose membranes. Sera derived from HDM allergic subjects and healthy controls were hybridized and signals were detected with nitro-blue tetrazolium and 5-bromo-4-chloro-3'-indolyl phosphate. Levels of HDM specific IgE: subject 1: >100 kU/L; subject 2: 3.97 kU/L; subject 3: 0.50 kU/L.
Figure 8. Experimental schedule for IP sensitization and the therapeutic procedure. BALB/c mice were IP-sensitized with 4 ug of HDM crude extract on days 0 and 7. From days 14 to 35 the mice received LNIT with NS 5ul/mouse/day, DN-Dp 3ug/5ul/mouse/day or DEX 10ug/5ul/mouse/day. IT challenge with HDM crude extract of 3ug/30ul was conducted on day 28 and day 35. Mice were sacrificed on day 37.
Figure 1. Effects of LNIT with DN-Dp on systemic immune responses. Serum concentrations of antigen-specific IgE (a) and IgG2a (b) were measured by ELISA. Values are expressed as mean±SEM of optical density (O.D.) at 450 nm of mice in each group. *P<0.05, compared to the denatured Der p and non-treatment groups.
Figure 2. Effects of LNIT with DN-Dp on Th cell-related cytokine expression. Th cell-related cytokine mRNA of lung tissue was measured by qPCR. IFN-g, IL-4, IL-10, and IL-17 were expressed as relative mRNA expression using the 2(-△△CT) calculation method. Data represent the mean relative mRNA expression (±SD ) from 3 mice in each group. *P <0.05, compared to the denatured Der p and non-treatment groups.
Figure 3. Effects of LNIT with DN-Dp on pulmonary function of mice. The allergen challenge was conducted with Der p crude extract. Airway hypersensitivity to methacholine was measured 30 min after the second allergen IT challenge. *P<0.05, compared to the denatured Der p and non-treatment groups.
Figure 4. Lung pathology after LNIT with DN-Dp. Lung tissues were obtained 2 d after the second IT exposure to Der p crude extract in Der p-sensitized mice and examined by H and E staining (magnification 200x).
Figure 5. Effects of LNIT with DN-Dp on proinflammatory cytokine expression. Proinflammatory cytokine mRNA of lung tissue was measured by qPCR. IL-1b, IL-6 and TNF-a were expressed as relative mRNA expression using the 2(-△△CT) calculation method. Data represent the mean relative mRNA expression (±SD) from 3 mice in each group. *P<0.05, compared to the denatured Der p and non-treatment groups.
Figure 6. IHC analysis of proinflammatory cytokine expression in lung tissues. Lung tissues were obtained 2 d after the second IT exposure to Der p crude extract in Der p-sensitized mice (magnification 200x).
