FIGURE 5.

14DM in T. brucei. a and b, immunodetection of 14DM in Tbb procyclic (PF) and bloodstream forms (BF). a, whole cell extracts of procyclic and mice-isolated bloodstream form trypanosomes were probed by Western blotting with mouse anti-Tbb14DM antiserum. Recombinant (r) Tbb14DM served as positive control. b, mice-isolated bloodstream form cell extracts (10 mg) were immunoprecipitated with rat anti-Tbb14DM antiserum. The immunoprecipitated material (IP) was analyzed by Western blotting after non-reducing electrophoresis with mouse polyclonal anti-Tbb14DM antiserum. Controls were performed in the absence of either cell extracts or rat antiserum. Tbb14DM served as positive control. c, suppression of Tbb infection in mice by oral administration of VNI (20 mg/kg) or clotrimazole (CLT, 50 and 100 mg/kg). Clotrimazole was selected among several commercially available antifungal azoles by testing their inhibitory potency in reconstituted Tbb14DM reaction in vitro due to the limited amount of VNI. The differences observed between the mean of treated and mock-treated mice are statistically significant as assessed by Student's t test (p < 0.01). Error bars indicate S.E.