Immune activation is a powerful strategy applied in the treatment of infectious diseases and cancers. While a number of natural carbohydrate conjugates and their derivatives or mimics have been proved to be able to activate the immune system, the complexity and heterogeneity of these molecules have posed daunting challenges on their further research and applications, as in many cases the difficulties in quality control cause serious safety concerns for clinical use. In this study, a series of glycopeptides have been designed, and synthesized utilizing solid-phase peptide synthesis (SPPS) strategy. Several synthesized peptides have showed self-assembling ability to produce stable and uniform nanoparticles, and one of which displayed high binding affinity and uptaking efficiency toward macrophages in vitro, as indicated by upregulated expression level of two immune co-stimulatory molecules CD40 and CD86, and two pro-inflammation cytokines interleukin-6 and interleukin-12. Moreover, vaccination of C57BL/6 mice with OVA model antigen in combination with the optimal glycopeptide elicits high titers of IgG antibody and promotes antigen specific memory response on the IFN-γ secretion. These results have demonstrated the utility of glycosylated nanoparticles as effective immunostimulant, and potential adjuvant that could be applied for vaccination.