The aim of this study was to design and develop stable nanoemulsion formulations containing IFN-γ to probe their use as an immunomodulating agent. The nanoemulsions comprising distilled water, triglycerides of capric acid/caprylic, sobitan-oleate (SP), polysorbate 80 (TW) and propylene glycol (PG) were prepared through ultra-homogenization and characterized regarding droplet size, polydispersity, surface charge, preliminary and accelerated physical stability, and rheological properties. Stable nanoemulsions were selected to incorporate nano doses of IFN-γ (100 ng.mL-1). The influence of IFN-γ incorporated nanoemulsions on functional activity of mononuclear cell for Escherichia coli enteropathogenic was analyzed through superoxide release, phagocytosis, microbicidal activity and intracellular calcium release. The optimized formulation, comprising aqueous and oily phase of 10 % and 80 %, respectively, and surfactant mix ratio (SP/TW/PG) of 3.5/5.5/1.0, remained stable in extreme conditions during 90 days, displaying oil-in-water characteristics, biocompatible pH value, significant maintenance of its rheological profile, hydrodynamic radius of 205 nm, zeta potential of -40 mV and average dose of IFN-γ 91 ng.mL-1. The developed formulation did not alter the MN cell viability rates. Increased the superoxide release, phagocytosis index and intracellular calcium release of MN cells of human blood. Our findings indicate that the produced formulation improved the immunomodulatory activity of the IFN-γ.