PbAWT (MR4), PbAmif- (Leiden Malaria Group) (15 (link)), or PbAWT-GFP-luciferase (MR4) parasites were cycled between Swiss Webster mice and Anopheles stephensi mosquitoes. For erythrocytic infection, cryopreserved stocks of infected red blood cells (iRBCs) were injected (106 iRBCs/mouse), and blood parasitemia was monitored by Giemsa-stained blood smears and flow cytometry (14 ). For the pre-erythrocytic stage infection, salivary gland sporozoites were extracted from infected mosquitoes on day 19 post-blood meal infection. WT or Cd74−/− C57BL/6J mice were infected by i.v. tail injection of 2000 PbAWT, PbAmif- or PbAWT-GFP-luciferase sporozoites, and blood patency was monitored beginning day 3 by blood smear and flow cytometry. Liver parasite burden was monitored at 48 h after infection using an IVIS imaging system (Caliper) or quantitative PCR (14 ).
For adoptive transfer, splenocytes were isolated six days after infection of WT or Cd74−/− mice infected with 106PbAWT iRBC, CD8 T cells were purified with anti-CD8 (Ly-2, Miltenyi Biotech) according to the manufacturer’s protocol. 1×107 cells were transferred, i.v. into recipient C57BL6/J Cd8−/− or Cd8−/−Cd74−/− mice and mice infected three days after with 106PbAWT iRBC.
For adoptive transfer, splenocytes were isolated six days after infection of WT or Cd74−/− mice infected with 106PbAWT iRBC, CD8 T cells were purified with anti-CD8 (Ly-2, Miltenyi Biotech) according to the manufacturer’s protocol. 1×107 cells were transferred, i.v. into recipient C57BL6/J Cd8−/− or Cd8−/−Cd74−/− mice and mice infected three days after with 106PbAWT iRBC.