Female swiss mice

Female Swiss mice (body weight (BW) ~20–24 g; Janvier Labs, France) were allocated randomly to groups of three animals and infected intraperitoneally (i.p.) with 10⁴T. b. brucei Squib 427 derived from a heavily infected donor mouse. Drinking water and food were available ad libitum throughout the experiment. Analogue 9 was formulated in 10% (v/v) polyethylene glycol 400 in water at 2 mg mL⁻¹ or in 5% (v/v) Tween-80 in water at 4 mg mL⁻¹ and was freshly prepared at every administration. Analogue 9 was administered orally b.i.d. for 5 days at 25, 12.5 and 6.25 mg kg⁻¹ or i.p. s.i.d. for 5 days at 10 mg kg⁻¹ (all groups n = 3). The reference drug suramin was formulated in phospahte-buffered saline (PBS) at 2.5 mg mL⁻¹ and administered s.i.d. i.p. for 5 days at 10 mg kg⁻¹ (n = 3). All treatments were initiated 30 min prior to the i.p. infection. Animals were observed for the occurrence of clinical or adverse effects during the course of the experiment and were weighed daily. Parasitaemia was determined by microscopic evaluation of tail vein blood samples at 4, 7, 10, 14 and 21 dpi (pre-set endpoint). As a test of cure, blood samples (250 µL) were collected from treated mice at 21 dpi and were sub-inoculated i.p. in naive Swiss mice (n = 3), followed by monitoring of parasitaemia as follow-up.

Female Swiss mice (6–8 weeks old, from Janvier Labs) were used for all routine parasite infections. Conditional genome editing was performed in the P. berghei (ANKA strain) PbDiCre line, obtained after integration of mCherry and DiCre expression cassettes at the dispensable p230p locus [19 (link)]. Two additional lines expressing RON4-mCherry (bioRxiv 2021.10.25.465731) and/or GFP [44 (link)] were used for immunoprecipitation and electron microscopy experiments, respectively. Parasites were maintained in mice through intraperitoneal injections of frozen parasite stocks. Anopheles stephensi mosquitoes were reared at 24°C with 80% humidity and permitted to feed on infected mice that were anaesthetized, using standard methods of mosquito infection as previously described [45 (link)]. Post feeding, P. berghei-infected mosquitoes were kept at 21°C and fed daily on a 10% sucrose solution.

Female Swiss mice (Janvier Labs), 6–8 weeks old and weighing ∼30 g were used for the septicemia model. Experiments were monitored in the ARCHE-BIOSIT animal lab in Rennes, and were performed in biological duplicates. We used groups of 5 mice for the mild septicemia model. Mice were infected intravenously by the tail vein with 200 μl of suspensions in 0.9% NaCl containing 3 × 10⁸ bacteria with either HG003, HG003 ΔisrR, or HG003 ΔisrR locus2::isrR⁺ strains. Mouse survival was monitored for 8 days, and the statistical significance of difference(s) between groups was evaluated using the Mantel-Cox test. A p value < 0.05 was considered as significant. All experimental protocols were approved by the Adaptive Therapeutics Animal Care and Use Committee (APAFiS #2123–2015100214568502v4).

Female Swiss mice, BALB/c mice, and female golden hamsters were purchased from Janvier (Le Genest Saint Isle, France). Food for laboratory rodents (Carfil, Arendonk, Belgium) and drinking water were available ad libitum. Before the start of the in vivo experiments, the hamsters were randomly allocated to experimental units of 3 animals each.