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. 2012 Dec 19;5(2):250–263. doi: 10.1002/emmm.201201677

Figure 4. Protective antigens are probably present in the immunizing sporozoites.

Figure 4

All experiments were repeated three times (n = 3) with five mice per group (A) or by three triplicates (B) at each time.

  1. 100 × 103 CD11c+-DC were sorted by FACS after co-culture with γ-spz or medium for 1.5 h, and then injected i.v. into recipient naïve mice. One week later, the recipient mice were challenged i.v. with 5 × 103 PyWT and the appearance of blood stage parasites was monitored three times a week by microscopic examination of Giemsa-stained blood smears from day 4 to day 14 post sporozoites infection. Each immunization group had a naïve control group of five mice (n = 5) immunized with PBS.
  2. Exo-erythrocytic forms (EEFs) were detected by fluorescence microscopy after double staining with anti-CSP and anti-HSP70, in the FACS-purified DC after co-culture with γ-spz sporozoites (or medium) for 0, 24 or 48 h after the co-culture of DC. The infectiousness of the γ-spz was assessed in control wells containing primary mouse hepaotcyte (Hep). Results are expressed as mean ± standard deviation (SD).
  3. Parasites were also detected by RT-nPCR after co-culture of γ-spz and DC, in the DC fraction purified by FACS immediately, 24 or 48 h after the co-culture. The control for sporozoite infectiousness was HepG2-A16-CD81, which are highly receptive to P. yoelii sporozoites (Yalaoui et al, 2008). RT− is a control for DNA contamination prepared in the absence of the reverse transcriptase. + is a positive control for the amplification reaction from genomic P. yoelii 265BY DNA.