Figure captions
Figure. 1 Effect of recombinant TSA-1-C4 plus Tc24-C4
combination on NO and H2O2 production by
peritoneal macrophages. Peritoneal cells isolated from naïve mice were
cultured in supplemented DMEM for 48 h, non-adherent cells were removed
through DMEM washing. Cells were stimulated with TSA-1-C4 and Tc24-C4
combination (100, 50, 25, 12.5 and 6.25 µg/ml) for 48 h. Supernatants
were recovered to measure NO (indirectly by nitrite concentration)(A) and H2O2 (B)production. Data is represented by the mean ± S.D. of three independent
experiments. Significance was calculated by one-way ANOVA and Tukey’s
multiple-comparison test (* recombinant protein stimulated
cells vs non stimulated cells; ^ Tc24-C4 stimulated cells vs
TSA-1-C4 stimulated cells; & Tc24-C4 stimulated cells vs
TSA-1-C4+Tc24-C4 stimulated cells). One, two, and three symbol
characters were used to annotate the P values of <0.05,
<0.01, and <0.001, respectively.
Figure. 2 Effect of recombinant TSA-1-C4 plus Tc24-C4
combination on cytokine production by peritoneal macrophages. Peritoneal
cells isolated from naïve mice were cultured in supplemented DMEM for 48
h, adherent cells were stimulated with TSA-1-C4 and Tc24-C4 combination
(100, 50, 25 and 12.5 and 6.25 µg/ml) for 48 h. Supernatants were
recovery to measure TNF-α (A), IL-1β (B), IL-6(C) and IL-10 (D) production. The purple dotted line
represents cytokine production by non-stimulated cells control. Data is
represented by the mean ± S.D. of three independent experiments.
Significance was calculated by one-way ANOVA and Tukey’s
multiple-comparison test (% TSA-1-C4+Tc24-C4 stimulated cells
vs TSA-1-C4 stimulated cells; & TSA-1-C4+Tc24-C4 stimulated
cells vs Tc24-C4 stimulated; ^ Tc24-C4 stimulated cells vs
TSA-1-C4 stimulated cells). One, two, and three symbol characters were
used to annotate the P values of <0.05,
<0.01, and <0.001, respectively.
Figure. 3 Effect of recombinant TSA-1-C4 plus Tc24-C4
combination on cytokine production by CD8+ T cells.
Peritoneal cells isolated from naïve mice were stimulated with TSA-1-C4
and Tc24-C4 combination (100, 50, 25 and 12.5 µg/ml); after 48 h,
supernatant was removed and cells were co-cultivated with
CD8+ T cells from naïve mice. Supernatants were
recovery to measure IFN-γ (A), TNF-α (B), IL-4(C) and IL-10 (D) production. The purple dotted line
represents cytokine production by non-stimulated cells control. Data is
represented by the mean ± S.D. of three independent experiments.
Significance was calculated by one-way ANOVA and Tukey’s
multiple-comparison test (% TSA-1-C4+Tc24-C4 stimulated cells
vs TSA-1-C4 stimulated cells; & TSA-1-C4+Tc24-C4 stimulated
cells vs Tc24-C4 stimulated). One, two, and three symbol characters were
used to annotate the P values of <0.05,
<0.01, and <0.001, respectively.
Figure. 4 (A ) Timeline for T.
cruzi -experimental infection using BALB/c mice, prime-boost vaccination
(red arrows), macrophages injection (purple arrow) and euthanize (blue
arrow). (B ) Effect of the recombinant protein combination onin-vivo cytotoxicity assay. Representative histogram profile
showed as percentage. Significance was calculated by one-way ANOVA and
Tukey’s multiple-comparison test and is indicated as follows ***,
P≤0.001 (comparing with the infected untreated control group).