Article Figures & Data
Figures
- FIG 1
In vitro parasiticidal effect of BIBW2992, CI-1033, and U0126 on E. multilocularis metacestode vesicle. Viability of vesicles was assessed upon treatment with 1 to 20 μM BIBW2992 (A), 1 to 40 μM CI-1033 (B), or 10 to 400 μM U0126 (C) for the indicated times. The LC50 (after 72 h of treatment) of each drug is shown.
- FIG 2
TEM images of E. multilocularis metacestode after in vitro treatment with BIBW2992, CI-1033, and U0126. Control vesicles treated with the solvent DMSO (A) or ethanol (EtOH) (B). (C) Enlarged view of the zone indicated in panel B. (D to F) Vesicles treated with BIBW2992 at concentrations of 0.5 μM for 72 h (D), 2 μM for 72 h (E), and 10 μM for 5 days (F). (G to I) Vesicles treated with 20 μM CI-1033 for 24 h (G), 48 h (H), and 72 h (I). (J to L) Vesicles treated with U0126 at concentrations of 100 μM for 24 h (J), 200 μM for 72 h (K), and 200 μM for 5 days (L). LL, laminated layer; GL, germinal layer; Teg, tegument; Uc, undifferentiated cells; Gly, glycogen storage cells; ld, lipid droplets. The white arrows in panels A to C indicate normal microtriches. White asterisks in panels D, H, and J indicate microtriches with a great reduction in number. Black arrows in panels E and I indicate cells with nuclear chromatin condensation and margination. The black asterisk in panel G indicates vacuolated cells. The black arrowheads in panel K indicate cells containing vacuoles and membrane stacks. Scale bar, 2 μm (for all figure panels).
- FIG 3
Assessment of proapoptotic effect of BIBW2992 on E. multilocularis metacestode vesicles in vitro. (A) DAPI staining images of vesicles treated with DMSO or 10 μM BIBW2992 (BIBW) for 48 h. Arrows indicate apoptotic cells. Scale bar, 20 μm. (B and C) Quantification of apoptotic cells in vesicles treated with 0.2 to 10 μM BIBW2992 for 48 h (B) or 10 μM BIBW2992 for 24 to 72 h (C). UT, untreated. (D) Caspase-3 activity of vesicles after exposure to 10 μM BIBW2992 for 48 h. Data in panels B to D are shown as means ± the standard deviations (SD). *, P < 0.05; ***, P < 0.001; N.S, not significant.
- FIG 4
Proapoptotic effect of BIBW2992 on E. multilocularis germinative cells. (A) Vesicles were administered to a 4-h EdU pulse to label proliferating germinative cells (red) and then treated with DMSO or 10 μM BIBW2992 for another 8 h. Arrows indicate apoptotic EdU+ cells. Scale bar, 10 μm. (B) Quantification of apoptotic EdU+ cells in DMSO- or BIBW2992-treated vesicles. The data are shown as means ± the SD.
- FIG 5
In vivo efficacies of BIBW2992 and U0126 against E. multilocularis in experimentally infected BALB/c mice. All treatments were performed 8 weeks postinfection and proceeded for 6 weeks. Each treatment group comprised 10 mice. (A) Mice were orally given 0.5% MC plus 0.5% CMC (vehicle control), albendazole (ABZ) in CMC (200 mg/kg/day), or BIBW2992 in MC (25 mg/kg/day 5 days a week). (B) Mice were given intraperitoneally 40% DMSO (vehicle control) or U0126 (25 μmol/kg three times per week). After euthanasia, parasite tissue was resected and weighed. Parasite weights are shown as box plots. *, P < 0.05; **, P < 0.01. (C to G) TEM images of E. multilocularis metacestodes obtained from vehicle-treated mice (C, MC + CMC; D, DMSO), albendazole-treated mice (E), U0126-treated mice (F), and BIBW2992-treated mice (G). LL, laminated layer; GL, germinal layer; H, host tissue. Scale bar, 2 μm.
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