Aim of the present study was to develop mannosylated niosomes as oral DNA vaccine carriers for the induction of humoral, cellular and mucosal immunity. Niosomes composed of span 60, cholesterol and stearylamine as constitutive lipids were prepared by reverse phase evaporation method and were coated with a modified polysaccharide o-palmitoyl mannan (OPM) in order to protect them from bile salt caused dissolution and enzymatic degradation in the gastrointestinal tract and to enhance their affinity towards the antigen presenting cells of Peyer's patches. Prepared niosomes were characterized in vitro for their size, shape, entrapment efficiency, ligand binding specificity and stability in simulated gastric fluid and simulated intestinal fluid. OPM coated niosomes were found to better stable in simulated GIT conditions. The immune stimulating activity was studied by measuring serum anti-HBsAg titer, secretory IgA level in intestinal and salivary secretions and cyokines level (IL-2 and IFN-gamma) in spleen homogenates following oral administration of niosomal formulations in Balb/c mice and compared with naked DNA as well as pure recombinant HBsAg injected intramuscularly. The serum anti-HBsAg titer obtained after oral administration of OPM coated niosomal formulations was although less as compared to that elicited by naked DNA and pure HBsAg administered intramuscularly, but the mice were seroprotective within 2 weeks and antibody level far above the clinically protective limit for humans was achieved. Intramuscular naked DNA and recombinant HBsAg did not elicited sIgA titer in mucosal secretions that was induced by oral administration of OPM coated niosomes. Similarly, cellular response (cytokines level) was absent in pure HBsAg treated animals. OPM coated niosomes produced humoral (both systemic and mucosal) and cellular immune response upon oral administration. The study signifies the potential of OPM coated niosomes as DNA vaccine carrier and adjuvant for effective oral immunization.