Figure 1
Replication of T. cruzi and T. brucei forms are inhibited by I-17. (A) T. cruzi epimastigotes were inoculated at 2 × 106 parasites per mL and incubated at 28 °C in LIT medium containing the indicated concentrations of NCPdCPU (an inactive 1,3-diarylurea, circles) or I-17 (squares). The numbers are means ± standard deviation (n = 3) of parasite counted after 48 hours of incubation. (B) T. brucei BSF (circles) and PCF (triangles) were inoculated at 2 × 105 parasites per mL in their respective culture media in the presence of the indicated concentrations of I-17, or NCPdCPU (closed symbols). The numbers are means ± standard deviation (n = 3) of parasites counted after 24 hours at 37 °C and 48 hours at 28 °C, respectively for BSF and PCF. (C) Relative viability of L6 myoblasts measured by activity of lactate dehydrogenase released after 72 hours in the presence of the indicated concentrations of I-17. (D) TCT was used to infect L6 cells. After infection the cells were treated for 72 hours with the indicated concentrations of I-17 and the number of intracellular amastigotes was determined by optical microscopy as described in Methods. The bars represent means ± standard deviation of three independent experiments. In all panels, *indicate p < 0.05, **p < 0.01 and ***p < 0.001 calculated using the Student’s T-test comparing to non-treated cells. (E) Serial dilutions of I-17 was used to treat U-2 OS cells infected with T. cruzi TCT. Compound activity is indicated as percentage of parasite free cells (black circles) and the host cell survival (empty squares), both calculated as described in Methods. (F) Calculation of EC50 values for T. cruzi and host cells expressed in µM are also explained in Methods. The values in (E) and (F) represent means ± standard deviation of three independent experiments.
