Objectives: This study aims to investigate the effects of cholesterol content and cationic property of liposomes on immune response. Materials and methods: Liposomes containing high amounts of 3β-[N-(N',N'-dimethylaminoethane)-carbamoyl] cholesterol hydrochloride (DC-cholesterol) were prepared and loaded with K- and D-type CpG oligonucleotide (CpG-ODN) via dehydration-rehydration (DRV) method. After splenocytes and peritoneal exudate cells (PECs) primed with lipopolysaccharide (LPS) was incubated either with free or liposomal CpG-ODN counterparts, supernatants were collected and used in cytokine (IFN-g, IL-1γ and IL-1β) ELISA. Additionally, supernatants of PECs primed with LPS and stimulated with liposomes containing different doses of DC-cholesterol were collected and used in IL-1β ELISA assay. Results: Low-dose CpG-ODN loaded liposomal formulations induced higher immune activation than free CpG-ODN at the same dose. While high-dose liposomal CpG-ODN formulations decreased pro-inflammatory cytokine production in splenocytes, they increased the secretion of IL-1β. Inflammasome activation was increased in a dose dependent manner when PECs primed with LPS were incubated with only liposomes. Varying lipid molar ratios of DC-Cholesterol containing liposomes increased IL-1β production based on increasing lipid molar ratio. Conclusion: This study revealed that type and lipid ratio of liposomes may alter the cellular efficacy of the loaded immune-stimulatory agent and may initiate inflammasome activation. © 2014 Turkish Journal of Immunology.