Example 48
In vivo adapted P. falciparum (3D7HLH/BRD) were selected as described in PLoS One 3, e2252 (2008). In brief, NSG mice (n=2 per experimental group; female; 4-5-week-old; 19-21 g; The Jackson Laboratory) were intraperitoneally injected with 1 ml of human erythrocytes (0-positive, 50% haematocrit, 50% RPMI 1640 with 5% albumax) daily to generate mice with humanized circulating erythrocytes (huRBC NSG). Approximately 2×107 blood-stage P. falciparum 3D7HLH/BRD (FASEB J. 25, 3583-3593 (2011)) were intravenously infected to huRBC NSG mice and >1% parasitaemia was achieved 5 weeks after infection. After three in vivo passages, the parasites were frozen and used experimentally. Approximately 48 h after infection with 1×107 blood-stage of P. falciparum 3D7HLH/BRD, the mean parasitaemia was approximately 0.4%. huRBC NSG mice were orally treated with a single dose of compound and parasitaemia was monitored for 30 days by IVIS to acquire the bioluminescence signal (150 mg kg-1 of luciferin was intraperitoneally injected approximately 10 min before imaging).
