Although most pathogens use the mucosal routes for invasion, the majority of currently available vaccines are administered parenterally. Injectable vaccines induce good systemic immunity but often unsatisfactory mucosal immunity. A non-injectable mucosal vaccine, which can be self-administered intranasally, may provide both effective systemic and mucosal immunity and can be used for vaccination of large populations within a short period of time in case of a sudden epidemic. Here, we report on a new intranasal (i.n.) influenza vaccine, based on a novel polycationic sphingolipid, N-palmitoyl D-erythro-sphingosyl carbamoyl-spermine (ceramide carbamoyl-spermine = CCS), having combined carrier and adjuvant activities, which elicits, in mice, strong systemic (serum) and local (lung and nasal) humoral and cellular responses, and provides protective immunity. In a comparative study, we show that both unmodified commercial vaccine and vaccine formulated with neutral or anionic liposomes were poorly immunogenic upon i.n. administration. Of five vaccine formulations based on well-established monocationic lipids in the form of unsized liposomes, three (DC-Chol, DDAB, and DSTAP-based) resulted in low serum and local responses, while two others (DMTAP and DOTAP-based vaccines) induced both systemic and local vigorous Th1+Th2 immune responses. However, only the vaccine formulated with CCS was equivalent or superior to the commercial vaccine co-administered with cholera toxin as an adjuvant. Furthermore, the CCS-based influenza vaccine was highly efficacious following a single or a repeated (x2) i.n. or a single i.m. administration, without an added adjuvant, in both young (2 months) and old (18 months) mice. It elicited high titers of strain cross-reactive hemagglutination inhibition (HI) antibodies, and the high antibody titers and protective immunity persisted for at least 9 months. No systemic adverse effects, and only a mild local inflammatory response, were observed in mice and rabbits vaccinated i.n. with the CCS vaccine formulation. A similar approach may prove efficacious for i.n. vaccination against other pathogens.