The injectable vaccines used today induce pathogen-specific immunoglobulin (Ig) G and T helper (Th) 1 responses to halt the progression of infectious diseases. However, most pathogens enter the body through the mucosa, and the induction of pathogen-specific mucosal immunity involving secretary Ig A (SIgA) and Th17 responses is essential for inhibiting their entry. Unfortunately, injectable vaccines are poor inducers of antigen (Ag)-specific mucosal immunity, because it is considered to be induced by unique mucosal dendritic cell (DC) subsets. Our previous studies showed that CD103+CD11b+ DCs in small intestinal lamina propria (siLPDCs) induce intestinal mucosal immunity by Toll-like receptor ligand stimulation via flagellin or CpG oligodeoxynucleotides (CpG-ODN). Unlike splenic DCs (SPDCs), they specifically express retinoic acid-converting enzyme retinal dehydrogenase isoform 2 (RALDH2), and can induce Ag-specific SIgA and Th17 responses in addition to IgG and Th1 responses. Here, we show that SPDCs co-stimulated by CpG-ODN and curdlan induce similar immune responses to those induced by activated siLPDCs. This adjuvant combination not only induces activation of SPDCs but also up-regulates their expression of RALDH2 and TGF-b, critical factors for IgA and Th17 induction. Intramuscular Ag injection with CpG-ODN and curdlan induces Ag-specific systemic responses (e.g., IgG and Th1 production). Surprisingly, the priming immunization also induces Ag-specific SIgA in stool, thereby protecting against cholera toxin-induced diarrhoea. Although the Ag-specific SIgA induction is transient after priming, oral Ag administration induces high levels of Ag-specific IgA in stools that exceeded 3 months. Moreover, Ag-specific Th1 and Th17 responses are induced in intestine after boosting. The boosting effect is also induced in the lungs by intranasal Ag administration, thereby effectively inhibiting colonization and invasion by Streptococcus pneumoniae. This prime-boost method provides an innovative injectable hybrid mucosal vaccine capable of effectively preventing pathogen invasion and disease progression in the targeted mucosal tissues.