Supplementary MaterialsData_Sheet_1. of TLR signaling in sensitization-recruited inflammatory monocytes attenuates OVA-induced allergic asthma by promoting the expression of Th1-associated cytokines. 0.05 was considered statistically significant. Results TLR signaling plays a protective role in OVA-induced allergic asthma To find out the cellular and molecular mechanisms of the protective function of TLR signaling in allergic asthma, we established the mouse model of OVA-induced allergic asthma by peritoneal sensitization using OVA/alum and intranasal challenge with OVA (Figures 1A,B), which is the most widely used mouse model of allergic asthma (24). To confirm if TLR signaling had a protective Bafetinib enzyme inhibitor role in our mouse model, we established OVA-induced allergic asthma in 0.05, ** 0.01, *** 0.0001, **** 0.00001, ns, not significant. Open in a separate window Figure 2 Activation of TLR signaling protects mice against OVA-induced allergic asthma. (ACE) 6-weeks-old WT mice were treated i.p., Bafetinib enzyme inhibitor with 10 g of Pam3CSK4 or 25 g of CpG 1 day before OVA/alum Bafetinib enzyme inhibitor sensitization and OVA challenge in the mouse model of OVA-induced allergic asthma. All mice were harvested and analyzed 1 week after OVA challenge (A). Histologic sections of lungs from each group were analyzed by H&E. Pictures show representative samples of 4C6 mice/group. Scale bar = 200 m (B). Histological scores for assessment of lung injury were shown (C). Total cells in BALF were counted (D), and the frequency and number of eosinophils in BALF were analyzed by FACS (E). Data are representative of three or more independent experiments with 4 mice per group. Data are the mean SEM. ** 0.01, **** 0.00001, ns, not significant. TLR signaling attenuates OVA-induced allergic asthma by affecting sensitization To identify which phase (e.g., sensitization or challenge) of OVA-induced allergic asthma could be affected by TLR signaling, we treated WT mice with Pam3CSK4 by peritoneal and intranasal routes, respectively (Figure ?(Figure3A).3A). We found that only treating mice i.p., with Pam3CSK4 protected against OVA-induced allergic asthma, characterized by reduced lung injury and less inflammatory cells, especially eosinophils, infiltration in alveoli when compared with WT control mice, but treating mice intranasally (i.n.) with Pam3CSK4 had no this protective effect (Figures 3BCE), implying that TLR agonist treatment might mainly influence the sensitization response in the peritoneal cavity. To prove this hypothesis, different from the mode of administration in Figure ?Figure3A,3A, we only treated mice i.p., with Pam3CSK4 at the sensitization stage, and the Pam3CSK4 was i.p., injected after OVA/alum sensitization (Figures ?(Figures3F).3F). The results showed that Pam3CSK4 injection only at sensitization stages showed equivalent protection compare to the mode of administration in Figures 3A,G. Thus, these results indicated that TLR signaling-mediated protective effect mainly affect the sensitization phase of OVA-induced allergic asthma. Open in a separate window Figure 3 TLR exerts protective function at sensitization phase of OVA-induced allergic asthma. (ACE) 6-weeks-old WT mice were treated i.p., or i.n., with 10 g of Pam3CSK4 1 day before OVA/alum sensitization and OVA challenge in the mouse model of OVA-induced allergic asthma. All mice were harvested and analyzed 1 week after OVA challenge (A). Histologic sections of lungs from each group were analyzed by H&E. Pictures Mouse monoclonal to CD13.COB10 reacts with CD13, 150 kDa aminopeptidase N (APN). CD13 is expressed on the surface of early committed progenitors and mature granulocytes and monocytes (GM-CFU), but not on lymphocytes, platelets or erythrocytes. It is also expressed on endothelial cells, epithelial cells, bone marrow stroma cells, and osteoclasts, as well as a small proportion of LGL lymphocytes. CD13 acts as a receptor for specific strains of RNA viruses and plays an important function in the interaction between human cytomegalovirus (CMV) and its target cells show representative samples of 4C6 Bafetinib enzyme inhibitor mice/group. Scale bar = 200.