By 15 days after infection, 12 mice had died in each control group (20% survival rate) and
11 in the subcutaneous immunization selleck screening library group (26.6% survival rate), this difference not being statistically significant (χ2= 0.186, P= 0.666). In the intranasal immunization group six mice had died (60% survival rate) (Fig. 4), this difference in survival rate being statistically significant (χ2= 5.000, P= 0.025). Therefore nasal immunization is more effective than subcutaneous immunization against EHEC O157:H7. In this study, we analyzed β-turn, flexibility, hydrophilicity, accessibility, antigenicity and other parameters of IntC300 using DNASTAR software and the protein network server from Harvard University. We found that peptides 658–669 (KASITEIKADKT), 711–723 (QTQATTGNDGRAT), 824–833 (KATSGDKQTV), 897–914 (KQTSSEQRSGVSSTYNLI) and 919–931 (LPGVNVNTPNVYA) were potential B-cell epitopes of intimin γ. There are nine shared amino acids (ITEIKADKT)
between the KT-12 peptide sequence (KASITEIKADKT) predicted in this study for EHEC O157:H7 IntC300 and that validated by Adu-Bobie et al. (ITEIKADKTTAVANGQDAIT), which is the peptide sequence for EPEC O126:H7 IntC280 (20). Since there is about 87% homology between EHEC and EPEC in the eae gene, it is likely that this gene has a similar function in both find protocol strains. Thus, there was a high possibility that KT-12 might serve as an antigenic site. This study showed that intranasal Amrubicin and subcutaneous immunization of KT-12-KLH conjugate both induce high concentrations of IgG antibodies. Nasal-mucosal immunization induced a high concentration of IgA antibodies, whereas subcutaneous immunization did not. The survival rate of the nasal immunization group was higher than that of the subcutaneous immunization group after infection of the animals with EHEC O157:H7. This suggests that while subcutaneous immunization can induce a higher concentration of IgG,
its protective effect is not strong enough to block infection with EHEC O157:H7, probably because such protection is mainly mediated through IgA and other antibodies, and not by IgG. EHEC O157:H7 invades the human body through the digestive tract, adhering only to the intestinal mucosa without invading epithelial cells. Epithelial cells can actively transport secretory IgA, but not IgG, antibodies (21). High concentrations of IgA can block infection at the primary stage, whereas IgG cannot. These factors may in part explain why intranasal immunization exerts better protection than subcutaneous immunization. Another important factor is the presence of the CMIS: mucosal immunization in one part of the body can induce mucosal immune response in distant parts of the body. Thus, antigen-specific B and T cells can migrate from nasal mucosa-associated lymphoid tissue to regional lymph nodes, enter the blood circulation, and finally reach their target sites.