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Background  Searching for new effective drugs against human and animal toxoplasmosis we decided to test the anti- Toxoplasma  potential of phytoecdysteroids (α-ecdysone and 20-hydroxyecdysone) characterized by the pleiotropic activity on mammalian organisms including the enhancement of host’s anti-parasitic defence. This objective was accomplished by the  in vitro  evaluation of  T. gondii  growth in phytoecdysteroid-treated immunocompetent cells of selected hosts: humans and two strains of inbred mice with genetically determined different susceptibility to toxoplasmosis.    Methods  Peripheral mononuclear blood cells were isolated from  Toxoplasma -positive and  Toxoplasma -negative women ( N  = 43) and men ( N  = 21). Non-infected mice (C57BL/6,  N  = 10 and BALB/c,  N  = 14) and mice (BALB/c,  N  = 10) challenged intraperitoneally with 5 tissue cysts of the  T. gondii  DX strain were also used in this study as a source of splenocytes. The effects of phytoecdysteroids on the viability of human PBMC and mouse splenocytes were evaluated using the MTT assay. The influence of phytoecdysteroids on PBMCs, splenocytes and  T. gondii  proliferation was measured using radioactivity tests (the level of 3[H] uracil incorporation by toxoplasms or 3[H] thymidine by PBMCs and splenocytes), which was confirmed by quantitative Real-Time PCR. Statistical analysis was performed using SigmaStat 3.5 (Systat Software GmbH). The best-fit IC50 curves were plotted using GraphPad Prism 6.0 (GraphPad Software, Inc.).    Results  Our results showed that phytoecdysteroids promote the multiplication of  Toxoplasma  in cultures of human or murine immune cells, in contrast to another apicomplexan parasite,  Babesia gibsoni . Additionally, the tested phytoecdysteroids did not stimulate the  in vitro  secretion of the essential protective cytokines (IFN-γ, IL-2 and IL-10), neither by human nor by murine immune cells involved in an effective intracellular killing of the parasite.    Conclusions  Judging by the effect of phytoecdysteroids on the  T. gondii  proliferation, demonstrated for the first time in this study, it seems that these compounds should not be taken into consideration as potential medications to treat toxoplasmosis. Phytoecdysteroids included in the food are most likely not harmful for human or animal health but certain nutrients containing ecdysteroids at high concentrations could promote  T. gondii  proliferation in chronically infected and immunocompromised individuals. In order to assess the real impact of ecdysteroids on the course of natural  T. gondii  invasion,  in vivo  research should be undertaken because it cannot be ruled out that the  in vivo  effect will be different than the  in vitro  one. However, taking into account the possible stimulating effect of ecdysteroids on some opportunistic parasites (such as  Toxoplasma  or  Strongyloides ) further studies are necessary and should focus on the mechanisms of their action, which directly or indirectly enhance the parasite growth. Since ecdysteroids are considered as potential drugs, it is essential to determine their effect on various parasitic pathogens, which may infect the host at the same time, especially in immunocompromised individuals.

Phytoecdysteroids as modulators of the Toxoplasma gondii growth rate in human and mouse cells