8% and 58.7 ± 4.9% of ExPEC strain PCN033 were killed with hyperimmune mouse sera against OmpC and OmpF, respectively. The results indicated that sera from both OmpC- and OmpF-immunized mice could mediate a significantly higher level of opsonophagocytic killing of ExPEC than sera from mice that received adjuvant

alone. The evidence for recombination signals was found in the E. coli ompC alignment by two programs, SBP and GARD, used for testing recombination. Staurosporine cell line Three potential recombination breakpoints with significant phylogenetic incongruence were identified at the nucleotide positions 492, 744 and 981 in the alignment of ompC. Four non-recombinant alignments together with the relevant trees of topological congruity were generated for the subsequent selection analysis. Based on the FEL inference, the selection profile of the ompC coding region is illustrated in Fig. 5a. The porin showed significant evidence for positive selection, with seven codon sites (47, 189, 223, 237, 322, 324 and 325) under positively selected force. Structural mapping of these sites is shown in Fig. 5b. According to the predicted topology of OmpC by pred-tmbb, all positively selected sites were located in the extracellular space and outer membrane surface. In addition, 48 negatively selected sites were detected at the 0.1 significance level. For ompF, two recombination

breakpoints were identified at the nucleotide positions 234 and 809 in the gene alignment. Notably, none of the positively selected sites was detected in the ompF gene through FEL inference. The E. coli genes ompC and ompF each encode an outer AZD0530 ic50 membrane protein. OmpC has a narrower pore and is preferentially expressed under higher osmolar pressure compared with OmpF (Nikaido, 2003). OmpC and OmpF have both been functionally confirmed to be beta barrel porins (Basle et al., 2006), which are important HAS1 for

dynamic interactions with the host immune system (Massari et al., 2003). Additionally, OmpC and OmpF are involved in antibiotic resistance and bacterial virulence (Negm & Pistole, 1999; De et al., 2001; Kumar et al., 2010). In this study, the immunogenic properties of porcine ExPEC OmpC and OmpF were investigated using a mouse model. Both porins OmpC and OmpF of ExPEC can provide high protection against lethal infection with the highly virulent strain PCN033. In addition, OmpC and OmpF both could induce high titers of IgG antibodies, indicating that these two proteins have good immunogenic properties. The type of immune responses was reflected by the two IgG subclasses produced through immunization, IgG1 and IgG2a. In mice, serum IgG1 is associated with a Th2-type response, whereas serum IgG2a is associated with a Th1-type response, which is particularly effective at mediating bacterial opsonophagocytosis (Unkeless et al., 1988). Our study showed that OmpC and OmpF elicited high titers of IgG2a, although less than IgG1, which indicated that OmpC and OmpF could induce significant Th1/Th2 immune responses.