Toll-like receptor 5 recognizes bacterial flagellin, plays a critical role in innate immunity, and contributes to flagellin-specific humoral immunity. Further, TLR5-expressing dendritic cells play an important role in IgA synthesis in the intestine; however, the contribution of TLR5 to antigen (Ag)-specific mucosal immunity remains unclear. Thus, whether TLR5 is essential for the induction of intestinal secretory (S)IgA antibody (Ab) responses against flagellin and bacterial Ags attached to the bacterial surface in response to an oral flagellated bacterium,Salmonella, was explored in this study. Our results indicate that when TLR5 knockout (TLR5−/−) mice are orally immunized with recombinantSalmonellaexpressing fragment C of tetanus toxin (rSalmonella-Tox C), tetanus toxoid (TT)- and flagellin (FliC)-specific systemic IgG and intestinal SIgA Abs are elicited. The numbers of TT-specific IgG Ab-forming cells (AFCs) in the spleen and IgA AFCs in the lamina propria (LP) of TLR5−/− mice were comparable to those in wild-type mice. rSalmonella-Tox C was equally disseminated in TLR5−/− mice, TLR5−/− mice lacking Peyer's patches (PPs), and wild-type mice. In contrast, TLR5−/− PP-null mice failed to induce TT- and FliC-specific SIgA Abs in the intestine and showed significantly reduced numbers of TT-specific IgA AFCs in the LP. These results suggest that TLR5 is dispensable for the induction of flagellin and surface Ag-specific systemic and mucosal immunity against oral flagellated bacteria. Rather, pathogen recognition, which occurs in PPs, is a prerequisite for the induction of mucosal immunity against flagellated bacteria.