Cellulose nanofibrils (CNFs), unique and promising natural materials have gained significant attention recently for biomedical applications, due to their special biomechanical characteristics, surface chemistry, good biocompatibility and low toxicity. However, their long bio-persistence within organisms may provoke chronic immune reactions and this aspect of CNFs has not been studied to date. Therefore, the aim of this work was to examine and compare the biocompatibility and immunomodulatory properties of CNFs in vitro. CNFs (diameters of 10-70nm; lengths of a few microns) were prepared from Norway spruce (Picea abies) by mechanical fibrillation and high pressure homogenisation. L929 cells, rat thymocytes or human peripheral blood mononuclear cells (PBMNCs) were cultivated with CNFs. None of the six concentrations of CNFs (31.25µg/ml – 1mg/ml) induced cytotoxicity and oxidative stress in the L929 cells, nor induced necrosis and apoptosis of the thymocytes and PBMNCs. Higher concentrations (250µg/ml – 1mg/ml) slightly inhibited the metabolic activities of the L929 cells as a consequence of inhibited proliferation. The same concentrations of CNFs suppressed the proliferation of PBMNCs to phytohemaglutinine, a T-cell mitogen, and the process was followed by down-regulation of interleukin-2 (IL-2) and interferon-γ (IFN-γ) production. The highest concentration of CNFs inhibited IL-17A but increased IL-10 and IL-6 production. The secretions of the inflammatory cytokines, IL-1β and the tumor necrosis factor-α (TNF-α) as well as Th2 cytokine (IL-4), remained unaltered. In conclusion, the results suggest that these CNFs are biocompatible, non-inflammatory and non-immunogenic nanomaterial. Higher concentrations seem to be tollerogenic to the immune system, a characteristic very desirable for implantable biomaterials.