In the present paper we focused on the potential role of human lactoferrin for the intracellular replication rate of T. gondii BK tachyzoites following our earlier observation that Toxoplasma gondii was able to bind human lactoferrin but not serum transferrin. The study was performed in vitro on human CaCo-2 epithelial cells and mouse L929 fibroblasts. We found that the multiplication of the parasite was inhibited by lactoferrin in both cell lines used. However, the direct cytotoxic effect on the parasite and the host cells was not observed. The intracellular growth of T. gondii was not affected when tachyzoites or host cells were only pre-coated with human lactoferrin. The results suggest that lactoferrin does not influence parasites penetration into host cells but could trigger unknown antiparasitic mechanisms in the infected cells.