Vitamins, minerals, and nanocurcumin play a substantial role in various nutraceutical/pharmaceutical formulations that are widely used in therapeutics, cosmetics, and dietary supplements. The current study aimed to investigate the comparative in vitro immunomodulatory effect of a novel nanocurcumin-based formulation with curcumin in LPS-induced cytokine expression, NK cells’ activity, and phagocytosis. The proinflammatory cytokines (TNF-α, IL-1β, and MIP-1α) and NK cells’ activity were measured in cell supernatants using ELISA assay; however, phagocytosis activity was performed using colorimetric analysis. The chemical characterization of novel nanocurcumin-based formulation using LC–MS (R t 19.02 min) and mass spectra analysis (m/z 369.04) confirmed the presence of the curcumin in highest peak concentration. MTT assay in three tested cell-lines showed that the formulation was found non-toxic at all the tested concentrations. The expression of TNF-α, IL-1β, and MIP-1α in splenocytes was significantly (p ≤ 0.001) inhibited. Besides, the NK cells’ activity and phagocytosis (macrophage) were increased significantly (p ≤ 0.001). Overall, the promising results of this study indicated the significant immunomodulatory effect of nanocurcumin-based formulation compared to the curcumin, which could be used against various inflammatory disorders such as allergy, asthma, autoimmune diseases, coeliac disease, inflammatory bowel disease, etc.