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. 2014 Jul 15;9(7):e102225. doi: 10.1371/journal.pone.0102225

Figure 1. Immunization of mice with Pyfabb/f− reduces PyCSP-Luc in vivo luminescence.

Figure 1

(A–B) Expression of luciferase (Luc) and PyCSP-Luc in the liver of mice immunized by HTVI. (A) Luciferase signal in live naïve BALB/c mice injected by HTVI with 25 µg of either phCMV-Luc plasmid DNA (top panel) or phCMV-PyCSP-Luc (bottom panel) and imaged 8 h later by IVIS, after injection of D-luciferin. The scale indicates radiance expressed as p/s/cm2/sr. (B) Kinetics of luciferase signal in live naïve BALB/c mice (shown as total flux p per second) during the course of the experiment. Data is representative of two individual experiments, with 3 mice each per group. (C–E) Reduction of luciferase signal in the liver of Pyfabb/f−-immunized mice upon HTVI challenge with PyCSP-Luc plasmid DNA (C) Luciferase signal in 3 representative live mice immunized twice with 50,000 Pyfabb/f− salivary gland sporozoites (right panel) or mock-immunized with salivary gland debris (left panel), challenged by HTVI with 25 µg of phCMV-Luc plasmid DNA (top panel) PyCSP-Luc (bottom panel) 30 days after the last immunization, imaged 7 d post challenge as described. Each group contained 4 to 5 mice. (D) Quantification of luciferase signal (shown as total flux p per second) from mice in the four groups described in part C. The data represents 4 to 5 individually analyzed mice in each group and correspond to mean ± SD; significant differences between the mean of the mock vs. Pyfabb/f−-immunized mice for each plasmid calculated using the Mann-Whitney test are indicated (** = p<0.01). (E) Inhibition of luciferase signal over the course of the experiment for mice challenged with plasmid DNA encoding for luciferase alone (Luc, red triangles) or PyCSP-Luc (blue squares), calculated as percentage reduction vs. the mock-immunization control). The data represents 4 to 5 individually analyzed mice in each group and correspond to mean ± SD; significant differences between the mean of Luc vs. PyCSP-Lus calculated using the Mann-Whitney test are indicated for each plasmid (* = p<0.005; ** = p<0.001).