FIG 4.
ECM occurrence in Δhmgb2 ANKA-infected C57BL/6 mice supplemented with recombinant PbHMGB2 protein. (A) Survival analysis of C57BL/6 mice infected i.p. with 105 Δhmgb2 ANKA pRBCs and thereafter injected from days 4 to 8 p.i. (dashed line) with either PbHMGB2 (25 mg/kg) or vehicle (protein buffer plus 50 ng LPS, which takes into account the residual LPS contamination of the recombinant protein) twice a day (every 12 h) (n = 10 for each experimental group; the experiment was done 3 times). C57BL/6 mice were also infected i.p. with 105 WT P. berghei ANKA pRBCs as an ECM positive control. Mice were monitored for ECM symptoms from d5 p.i. (B) Parasitemia was measured by flow cytometry. Values represent means ± SD for one representative experiment of three. (C) The amount of Evans blue dye (BE) that infiltrated the brain was measured as described in Materials and Methods for the three sets of mice (n = 5 for each experimental group). Multiple comparisons were analyzed by the Kruskal-Wallis test followed by Dunn's posttest for analyses of differences in survival (P = 0.0025) and BBB permeability (P = 0.0008) between all sets of mice.