The IFN-γ pathway is central for ECM development after blood-stage PbA infection. We first assessed the role of this pathway

in preerythrocytic/intrahepatic stage infection by investigating ECM neurological signs development in IFN-γR1−/− mice. Following the injection of 1000 sporozoites, 60% of the WT control mice developed typical ECM neurological symptoms, such as ataxia, loss of grip strength, progressive paralysis, and coma, and succumbed within 8–9 days, as previously described [22]. In contrast, IFN-γR1−/− mice were fully resistant to the same challenge, surviving 30 days with no ECM neurological signs (Fig. 1A). Therefore, type II IFN-γ pathway is essential for ECM development after PbA sporozoite infection. The role of type I IFN-α/β versus type II IFN-γ pathways in ECM development after infection with hepatic or blood-stage PbA was then assessed in mice deficient for either IFNAR1 or IFN-γR1. RO4929097 datasheet LEE011 order IFNAR1−/− mice were partially protected against ECM following sporozoite-initiated infection, only 20% dying before day 10, and 40% eventually developing typical ECM neurological symptoms, which reflected a delayed ECM development after infection with sporozoites (Fig. 1A),

as compared with WT control mice, 60% of which developed ECM and died before day 10, and IFN-γR1-deficient mice, which were fully resistant to PbA challenge. After injection of PbA-parasited red blood cells (105 pRBC/mouse), WT mice succumbed within 7–9 days with typical ECM neurological signs,

while IFN-γR1−/− mice were resistant, surviving over 20 days after infection with no ECM neurological signs. IFNAR1−/− mice were partially protected, 41% dying before day 9 postinfection and a further 36% developing delayed ECM from day 9 to 11 (Fig. 1B). Partial protection of IFNAR1−/− mice was also seen in response to higher dose PbA-infected erythrocytes injection (106pRBC/mouse; data not shown). Parasitemia was analyzed by flow cytometry using GFP transfected parasites [23]. There was no delay in parasitemia in IFN-γR1−/− and IFNAR1−/− mice following either sporozoite or blood-stage PbA infection. At 9 days after Ergoloid PbA sporozoite infection, parasitemia was about 2% in all groups with no significant differences between WT, IFN-γR1−/−, and IFNAR1−/− mice (Fig. 1C), while after blood-stage PbA infection parasitemia was 11–12% at 7 days in WT and IFN-γR1−/− mice, and was slightly increased in IFNAR1−/− mice (Fig. 1D). IFN-γR1−/− and IFNAR1−/− mice succumbed at later stages to either sporozoite or blood-stage infection with high parasitemia (Fig. 1E and F) and severe anemia (Fig. 2A and B) in the absence of neurological signs. Thus, our data confirm the essential role of type II IFN-γ pathway in ECM development after either PbA merozoite or sporozoite infection and demonstrate a contribution of type I IFN-α/β pathways in ECM development that was not associated with any direct effect on parasite growth.